Team:KIT-Kyoto/nakagawa

From 2011.igem.org

(Difference between revisions)

Latest revision as of 02:41, 30 September 2011

8/25
（目的）
LBプレートの作成　PCR フェノクロ処理、電気泳動、制限酵素処理

（方法）
LBプレート作成

LB培地
bacto-agar	15 g/l

↓LB培地を作製する
↓1 Lあたり15 gのbacto-agarを入れて、オートクレーブ滅菌した
↓オートクレーブから取り出し、冷めないうちにスターラーで均一になるまで撹拌した
↓65°C程度に冷めたら、必要であれば抗生物質を加えた
↓10 cmのディスポプラスチックシャーレ1枚につき、20～25 ml分注した
↓水平な所に置いて固まらせた

PCR(GFP)

PCR条件

10 µM Primer F	1.5 µl
10 µM Primer R	1.5 µl
Template DNA	1 µl
10 x PCR Buffer for KOD Plus	5 µl
dNTPs	4 µl
MgSO₄	4 µl
ddH₂O	32 µl
KOD Plus	1 µl
	total 50 µl

Cycle条件

Pre-Denature	95°C	30sec
Denature	95°C	30sec	30 Cyle
Anneling	(60)°C	1min
Extension	68°C	1kb/min
End	4°C	keep

フェノクロ処理 ↓前回泳動した残りサンプルを合わせて135μlにした
↓ddH₂O65μl加えて全量を200μlにした
↓フェノール/クロロホルムを等量入れた
↓しっかりvoltexして、15000rpm,5分,25℃で遠心した
↓水層を別のエッペンに入れた（下の層がフェノール層）
↓次にクロロホルムのみで同様におこない、水層を別のエッペンに移した
↓1/10等量の3M酢酸ナトリウムを加えた
↓等量のイソプロパノールを加え、よく振った
↓12000rpm,20分25℃で遠心した
↓沈殿をとらないように上澄みだけ除いた
↓70%エタノールを200μl加えた
↓軽く混ぜた後12000rpm,10分,25℃で遠心
↓上澄みを除いた後乾燥させて長期保存ならTE、すぐに使うのでddH₂O50μlに溶かした

電気泳動
↓3本のＤＮＡ溶液サンプル　5μlにddH₂O0.3ml EcoR1　0.5μlPst1　0.5μl10×Buffer 0.7μlを加え37℃30分インキュベート
↓１%agaゲル20μlより0.2gのagaを使いそこにTMNを20ml加えてゲルとした
↓電子レンジで沸騰させて粒がなくなった後ゲルが固まるまで待った
↓1kbaseのマーカー5μlとDNAのLoading dye(×6)1μlとサンプル5μlとを足したもので計七個のサンプルをゲルの穴に注いだ
↓電気泳動　100V 30分
↓EtBr染色(手袋をつけて)　10分
↓紫外線ランプを付けた元で撮影

制限酵素処理

先ほどのDNA溶液50μlと切りたい場所の制限酵素（今回はPst1,EcoR1）を1μl、Bufferを使用する制限酵素にあわせたものを6μl（今回は10×H Buffer）、ddH₂Oを2μlを混ぜたものを37℃でインキュベートした 8/26
(目的)
大腸菌のベクターのトランスフォーム

(方法)
↓氷上でコンピテント細胞(DH5 Alpha:大腸菌株)を解凍した
↓前もって冷やしておいた1.5 mlチューブに100 µlのコンピテント細胞を分注した
↓余ったコンピテント細胞は-80°Cの冷凍庫に戻した
↓DNAをチューブに1～5 µl加えて、氷上で30分間冷やした
↓42°Cで45秒間熱ショックを与えた
↓0.9 mlのSOC培地を加えた
↓37°Cで1時間、振りながら回復培養した
↓1 mlをLBプレート(+amp,+kan,+camのいずれか)にまいた
↓37°Cで一晩培養した

(結果)
赤色のコロニーが多く生えてきた。GFPの色が赤色のDNAより大きく問題はなさそうであった。
8/29
(目的)
大会提出用ベクターPsb1C3の培養とアルカリミニプレップ
(方法)
8/26に培養した提出用ベクターのコロニーの生えたプレートを用いた
↓LBプレート(amp+)に大腸菌をまき、37°C(API2-MALT1は30°C)で一晩培養した
↓プレートからシングルコロニーを分離した
↓2 mlのLB培地で37°C(API2-MALT1は30°C)で一晩振とう培養した

(結果)
翌日コンタミしてると考えられるビン3本培養に成功したビンが3本できた
ただ念を入れて翌日同じ操作を行うことにした。

8/30
(目的)
大会提出用ベクターpsb1c3の培養→トラフォ(二回目) (方法)
8/26に培養した提出用ベクターのコロニーの生えたプレートを昨日同様用いた
↓LBプレート(amp+)に大腸菌をまき、37°C(API2-MALT1は30°C)で一晩培養した
↓プレートからシングルコロニーを分離した
↓2 mlのLB培地で37°C(API2-MALT1は30°C)で一晩振とう培養した

(結果)
翌日6本培養したがコンタミすることなく無事に大腸菌を回収することができた

8/31
(目的)
培養した菌液のアルカリミニプレップ、フェノールクロロホルム処理、電気泳動、制限酵素処理
アルカリミニプレを最初に行った。

Solution I	50 mM グルコース (MW 180)
	10 mM EDTA(pH 8.0)
	25 mM Tris-HCl (pH 8.0)
Solution II	0.2 N NaOH
	1% SDS
Solution III	3 M 酢酸カリウム
	1.8 M 酢酸

↓LBプレート(amp+)に大腸菌をまき、37°C(API2-MALT1は30°C)で一晩培養した
↓プレートからシングルコロニーを分離した
↓2 mlのLB培地で37°C(API2-MALT1は30°C)で一晩振とう培養した
↓1.5 mlの培養液を1.5 mlチューブにうつし、15,000 rpm、4°Cで5分間遠心し、上清を捨てた
↓100 µlの氷冷したSolution Iを加え、ボルテックスした
↓沈殿物を溶かした後、200 µlのSolution IIを加え、混ぜた、ボルテックスは使用しない
↓氷上で5分間冷やした
↓150 µlのSolution IIIを加え、混ぜた、ボルテックスは使用しない
↓氷上で5分間冷やした
↓15,000 rpm、4°Cで10分間遠心した
↓上清を新しいチューブにとり、沈殿物は廃棄した
↓450 µlのイソプロパノールを加え、混ぜた
↓2分間室温で放置した
↓15,000 rpm、室温で10分間遠心し、上清を捨てた
↓150～200 µlの70%エタノールを加え、軽くボルテックスした
↓2分間室温で放置した
↓15,000 rpm、室温で10分間遠心し、上清を捨てた
↓沈殿を10分～15分乾かした
↓RNaseのはいったTEを20 µl加えて37°Cで培養した

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-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/22</font></span></p>
+/25<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">トランスフォーメーション</span></p>
+（目的）<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
+LBプレートの作成　PCR フェノクロ処理、電気泳動、制限酵素処理<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓氷上でコンピテント細胞（</span><span lang="EN-US"><font face="Century">DH5 Alpha </font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">大腸菌体）解凍</span></p>
+<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓前もって冷やしておいた</span><span lang="EN-US"><font face="Century">1.5ml</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">チューブに</span><span lang="EN-US"><font face="Century">100</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">のコンピテント細胞を分注する</span></p>
+（方法）<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓</span><span lang="EN-US"><font face="Century">DNA</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">をチューブに</span><span lang="EN-US"><font face="Century">1~5</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">加えて氷上で</span><span lang="EN-US"><font face="Century">30</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分正確に冷やす</span></p>
+LBプレート作成<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓</span><span lang="EN-US"><font face="Century">42</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で</span><span lang="EN-US"><font face="Century">45</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">秒熱ショックを与える</span></p>
+<tr><td><table border=1 width="170px">
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓</span><span lang="EN-US"><font face="Century">0.1ml</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">を</span><span lang="EN-US"><font face="Century">LB</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">プレート（</span><span lang="EN-US"><font face="Century">+amp</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">）にまく</span></p>
+<tr><td width="100px" align=center>LB培地</td><td width="70px" align=right>&nbsp;</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓</span><span lang="EN-US"><font face="Century">37</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">度で一晩培養</span></p>
+<tr><td align=center>bacto-agar</td><td align=right>15 g/l</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
+</table>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/23</font></span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">PCR</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">プライマー設計　プレカルチャー</span></p>
+<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">大腸菌のベクターには</span><span lang="EN-US"><font face="Century">EXSP</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の制限酵素で切れる場所があり</span><span lang="EN-US"><font face="Century">EX DNA SP</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">となる</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">今回はまず</span><span lang="EN-US"><font face="Century">E</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">が</span><span lang="EN-US"><font face="Century">2~3X</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">が</span><span lang="EN-US"><font face="Century">2~3</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">で</span><span lang="EN-US"><font face="Century">S3</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">で</span><span lang="EN-US"><font face="Century">P</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">何塩基でもつけてよい。そして</span><span lang="EN-US"><font face="Century">GFP</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">につなげる上で</span><span lang="EN-US"><font face="Century">15</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">塩基必要なのでその条件で設計する。</span></p>
+↓LB培地を作製する<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">またプライマー設計においてもう片方はアンチコドンで設計する必要がある。</span></p>
+↓1 Lあたり15 gのbacto-agarを入れて、オートクレーブ滅菌した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">そしてプライマー作成サイトでは温度が</span><span lang="EN-US"><font face="Century">65</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">度を超えないように注意する</span></p>
+↓オートクレーブから取り出し、冷めないうちにスターラーで均一になるまで撹拌した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
+↓65°C程度に冷めたら、必要であれば抗生物質を加えた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">培養がうまくいっていたのでコロニーにマジックで印をつけて先を焦がしたつまようじで突っつくこれを</span><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">本分し、</span><span lang="EN-US"><font face="Century">2ml</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">ずつ</span><span lang="EN-US"><font face="Century">LB</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の液体培地に入れて</span><span lang="EN-US"><font face="Century">37</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で一晩培養（</span><span lang="EN-US"><font face="Century">+amp</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">）</span></p>
+↓10 cmのディスポプラスチックシャーレ1枚につき、20～25 ml分注した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
+↓水平な所に置いて固まらせた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/24</font></span></p>
+<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">アルカリミニプレッツ（プラスミド回収）</span></p>
+PCR(GFP)<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
+<table border="0"><tr><td>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">昨日の大腸菌が増えた菌液を取り出し</span><span lang="EN-US"><font face="Century">1.5ml</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">チューブに</span><span lang="EN-US"><font face="Century">1.4ml</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">程度入れる。その後</span><span lang="EN-US"><font face="Century">15000pm,5min,4</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で遠心</span></p>
+<table border="0" width="150px">
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">上澄みを廃棄し</span><span lang="EN-US"><font face="Century">glucose solution(sol1)</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">を</span><span lang="EN-US"><font face="Century">100</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μｌ加えて</span><span lang="EN-US"><font face="Century">voltex</font></span></p>
+<tr><td align=center>PCR条件</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">沈殿物を溶かした後</span><span lang="EN-US"><font face="Century">0.2NNaCl1%SDS200</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μｌ加えてよく混ぜる</span></p>
+</table>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">その後氷上</span><span lang="EN-US"><font face="Century">5</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分</span></p>
+<table border=1 width="220px">
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">氷冷した</span><span lang="EN-US"><font face="Century">3M</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の酢酸カリウムをいれて氷上五分</span></p>
+<tr><td width="150px" align=center>10 µM Primer F</td><td width="70px"  align=right>1.5 µl</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">15000rpm10min4</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で遠心</span></p>
+<tr><td align=center>10 µM Primer R</td><td align=right>1.5 µl</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">上澄みを新しいチューブに移す　入れて</span><span lang="EN-US"><font face="Century">voltex</font></span></p>
+<tr><td align=center>Template DNA</td><td align=right>1 µl</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">900</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の</span><span lang="EN-US"><font face="Century">2</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">‐プロパノールを加え混ぜて、二分待つ</span></p>
+<tr><td align=center>10 x PCR Buffer for KOD Plus</td><td align=right>5 µl</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">15000rpm,4</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃</span><span lang="EN-US"><font face="Century">,10</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分で遠心し、上澄みを捨てる</span></p>
+<tr><td align=center>dNTPs</td><td align=right>4 µl</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">１</span><span lang="EN-US"><font face="Century">ml</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の</span><span lang="EN-US"><font face="Century">70%</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">エタノールを加え</span><span lang="EN-US"><font face="Century">15000rpm,2</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分</span><span lang="EN-US"><font face="Century">,4</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で遠心する</span></p>
+<tr><td align=center>MgSO<sub>4</sub></td><td align=right>4 µl</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">沈殿物を乾かす</span></p>
+<tr><td align=center>ddH<sub>2</sub>O</td><td align=right>32 µl</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">電気泳動</span></p>
+<tr><td align=center>KOD Plus</td><td align=right>1 µl</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
+<tr><td>&nbsp;</td><td align=right>total 50 µl</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">本の乾燥ＤＮＡに</span><span lang="EN-US"><font face="Century">williQ0.3ml DNA5</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μｌ</span><span lang="EN-US"><font face="Century">EcoR10.5</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μｌ</span><span lang="EN-US"><font face="Century">Pst10.5</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μｌ</span><span lang="EN-US"><font face="Century">10</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">×</span><span lang="EN-US"><font face="Century">Buffer 0.7</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μｌ</span></p>
+</table>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">加え</span><span lang="EN-US"><font face="Century">37</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃</span><span lang="EN-US"><font face="Century">30</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分インキュベート</span></p>
+</td><TD></TD><TD></TD><td>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">１</span><span lang="EN-US"><font face="Century">%aga</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">ゲル</span><span lang="EN-US"><font face="Century">20</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">より</span><span lang="EN-US"><font face="Century">0.2g</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の</span><span lang="EN-US"><font face="Century">aga</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">を使いそこに</span><span lang="EN-US"><font face="Century">TMN</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">を</span><span lang="EN-US"><font face="Century">20ml</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">加えてゲルとする</span></p>
+<table border="0" width="100px">
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">電子レンジで沸騰させて粒がなくなった後約</span><span lang="EN-US"><font face="Century">20</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分待つ</span></p>
+<tr><td align=center>Cycle条件</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">1kbase</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">のマーカー</span><span lang="EN-US"><font face="Century">5</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">と</span><span lang="EN-US"><font face="Century">DNA</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">のマーカー</span><span lang="EN-US"><font face="Century">(</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">×</span><span lang="EN-US"><font face="Century">6)1</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">とサンプル</span><span lang="EN-US"><font face="Century">5</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">とを足したもので計七個のサンプルをゲルの穴に注ぐ</span></p>
+</table>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">電気泳動　</span><span lang="EN-US"><font face="Century">100V 30</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分</span></p>
+<table border=1 width="400px">
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">EtBr</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">染色</span><span lang="EN-US"><font face="Century">(</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">手袋をつけて</span><span lang="EN-US"><font face="Century">)</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">　</span><span lang="EN-US"><font face="Century">10</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分</span></p>
+<tr><td width="100px" align=center>Pre-Denature</td><td width="100px"  align=center>95°C</td><td width="100px"  align=center>30sec</td></td><td width="100px">&nbsp;</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">撮影</span></p>
+<tr><td align=center>Denature</td><td align=center>95°C</td><td align=center>30sec</td><td align=center rowspan=3>30 Cyle</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
+<tr><td align=center>Anneling</td><td align=center>(60)°C</td><td align=center>1min</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/25</font></span></p>
+<tr><td align=center>Extension</td><td align=center>68°C</td><td align=center>1kb/min</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">LB</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">プレートの作成　</span><span lang="EN-US"><font face="Century">PCR </font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">フェノクロ処理、電気泳動、制限酵素処理</span></p>
+<tr><td align=center>End</td><td align=center>4°C</td><td align=center>keep</td><td width="100px">&nbsp;</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
+</table>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">PCR</font></span></p>
+</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
+</table>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">フェノクロ処理</span></p>
+<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">前回泳動した残りサンプルを合わせて</span><span lang="EN-US"><font face="Century">135</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">にする</span></p>
+フェノクロ処理
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">milliQ65</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">加えて全量を</span><span lang="EN-US"><font face="Century">200</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">にする</span></p>
+↓前回泳動した残りサンプルを合わせて135μlにした<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">フェノール</span><span lang="EN-US"><font face="Century">/</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">クロロホルムを等量入れる</span></p>
+↓ddH<sub>2</sub>O65μl加えて全量を200μlにした<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">しっかり</span><span lang="EN-US"><font face="Century">voltex</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">して、</span><span lang="EN-US"><font face="Century">15000rpm,5</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分</span><span lang="EN-US"><font face="Century">,25</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で遠心する</span></p>
+↓フェノール/クロロホルムを等量入れた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">水層を別のエッペンに入れる（下の層がフェノール層）</span></p>
+↓しっかりvoltexして、15000rpm,5分,25℃で遠心した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">次にクロロホルムのみで同様におこない、水層を別のエッペンに移す</span></p>
+↓水層を別のエッペンに入れた（下の層がフェノール層）<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">1/10</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">等量の</span><span lang="EN-US"><font face="Century">3M</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">酢酸ナトリウムを加える</span></p>
+↓次にクロロホルムのみで同様におこない、水層を別のエッペンに移した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">等量のイソプロパノールを加え、よく振る</span></p>
+↓1/10等量の3M酢酸ナトリウムを加えた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">12000rpm,20</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分</span><span lang="EN-US"><font face="Century">25</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で遠心する</span></p>
+↓等量のイソプロパノールを加え、よく振った<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">沈殿をとらないように上澄みだけ除く</span></p>
+↓12000rpm,20分25℃で遠心した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">70%</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">エタノールを</span><span lang="EN-US"><font face="Century">200</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">加える</span></p>
+↓沈殿をとらないように上澄みだけ除いた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">軽く混ぜた後</span><span lang="EN-US"><font face="Century">12000rpm,10</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分</span><span lang="EN-US"><font face="Century">,25</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で遠心</span></p>
+↓70%エタノールを200μl加えた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">上澄みを除いた後乾燥させて長期保存なら</span><span lang="EN-US"><font face="Century">TE</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">、すぐに使うなら</span><span lang="EN-US"><font face="Century">milliQ50</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">に溶かす</span></p>
+↓軽く混ぜた後12000rpm,10分,25℃で遠心<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
+↓上澄みを除いた後乾燥させて長期保存ならTE、すぐに使うのでddH<sub>2</sub>O50μlに溶かした<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">制限酵素処理</span></p>
+<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">先ほどの</span><span lang="EN-US"><font face="Century">DNA</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">溶液</span><span lang="EN-US"><font face="Century">50</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">と切りたい場所の制限酵素（今回は</span><span lang="EN-US"><font face="Century">Pst1,EcoR1</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">）を</span><span lang="EN-US"><font face="Century">1</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">、</span><span lang="EN-US"><font face="Century">Buffer</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">を使用する制限酵素にあわせたものを</span><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">（今回は</span><span lang="EN-US"><font face="Century">10</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">×</span><span lang="EN-US"><font face="Century">H Buffer</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">）、</span><span lang="EN-US"><font face="Century">milliQ2</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">を混ぜたものを</span><span lang="EN-US"><font face="Century">37</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃でインキュベートする</span></p>
+電気泳動<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/26</font></span></p>
+↓3本のＤＮＡ溶液サンプル　5μlにddH<sub>2</sub>O0.3ml EcoR1　0.5μlPst1　0.5μl10×Buffer 0.7μlを
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">GFP</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">ゲル抽出</span></p>
+加え37℃30分インキュベート<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/28</font></span></p>
+↓１%agaゲル20μlより0.2gのagaを使いそこにTMNを20ml加えてゲルとした<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">大腸菌のコンピテント細胞に提出用ベクター</span><span lang="EN-US"><font face="Century">Psb1c3</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">を入れてインキュベート</span></p>
+↓電子レンジで沸騰させて粒がなくなった後ゲルが固まるまで待った<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">材料</span></p>
+↓1kbaseのマーカー5μlとDNAのLoading dye(×6)1μlとサンプル5μlとを足したもので計七個のサンプルをゲルの穴に注いだ<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">大腸菌のコンピテント細胞　</span><span lang="EN-US"><font face="Century">100</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span></p>
+↓電気泳動　100V 30分<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">クロラムフェ二コール　　　</span><span lang="EN-US"><font face="Century">10</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span></p>
+↓EtBr染色(手袋をつけて)　10分<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">LB</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">プレート</span></p>
+↓紫外線ランプを付けた元で撮影<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">大会提出用ベクター</span><span lang="EN-US"><font face="Century">pSB1C3 1</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span></p>
+<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">方法</span></p>
+制限酵素処理<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓提出用ベクター</span><span lang="EN-US"><font face="Century">pSB1C3</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の培養</span></p>
+<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓クリーンベンチにて大腸菌のコンピテント細胞を</span><span lang="EN-US"><font face="Century">1.5ml</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">のチューブに取る</span></p>
+先ほどのDNA溶液50μlと切りたい場所の制限酵素（今回はPst1,EcoR1）を1μl、Bufferを使用する制限酵素にあわせたものを6μl（今回は10×H Buffer）、ddH<sub>2</sub>Oを2μlを混ぜたものを37℃でインキュベートした
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓そこにクロラムフェ二コールを</span><span lang="EN-US"><font face="Century">10</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">と</span><span lang="EN-US"><font face="Century">pSB1C3</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">を</span><span lang="EN-US"><font face="Century">1</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">加える</span></p>
+/26<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓それを</span><span lang="EN-US"><font face="Century">LB</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">プレートにまき、ガラス棒でのばす</span></p>
+(目的)<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓</span><span lang="EN-US"><font face="Century">37</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で一晩インキュベートする</span></p>
+大腸菌のベクターのトランスフォーム<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">結果</span></p>
+<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">小さいながらもコロニーを確認することが出来た</span></p>
+(方法)<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/29</font></span></p>
+↓氷上でコンピテント細胞(DH5 Alpha:大腸菌株)を解凍した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">大会提出用ベクター</span><span lang="EN-US"><font face="Century">psb1c3</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の培養→トラフォ</span><span lang="EN-US"><font face="Century">(</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">一回目</span><span lang="EN-US"><font face="Century">)</font></span></p>
+↓前もって冷やしておいた1.5 mlチューブに100 µlのコンピテント細胞を分注した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">材料</span></p>
+↓余ったコンピテント細胞は-80°Cの冷凍庫に戻した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">スクリュービン</span><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">本</span></p>
+↓DNAをチューブに1～5 µl加えて、氷上で30分間冷やした<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">つまようじ</span><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">本</span></p>
+↓42°Cで45秒間熱ショックを与えた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/29</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">のコンピテント細胞を増やした</span><span lang="EN-US"><font face="Century">LB</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">培地</span></p>
+↓0.9 mlのSOC培地を加えた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">LB</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">液体培地　</span><span lang="EN-US"><font face="Century">12ml</font></span></p>
+↓37°Cで1時間、振りながら回復培養した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">クロラムフェ二コール　</span><span lang="EN-US"><font face="Century">12</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span></p>
+↓1 mlをLBプレート(+amp,+kan,+camのいずれか)にまいた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">方法</span></p>
+↓37°Cで一晩培養した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓クリーンベンチにて昨日のプレートにマジックでコロニーに</span><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">つ印を入れる</span></p>
+<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓スクリュービン一本に液体培地</span><span lang="EN-US"><font face="Century">2ml</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">、クロラムフェ二コール</span><span lang="EN-US"><font face="Century">2</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">加えたものを</span><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">本作る</span></p>
+(結果)<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓火であぶったつまようじで先ほど印をつけたコロニーをつつきスクリュービンに入れるこれを</span><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">本分行う</span></p>
+赤色のコロニーが多く生えてきた。GFPの色が赤色のDNAより大きく問題はなさそうであった。
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓その後</span><span lang="EN-US"><font face="Century">37</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で一晩インキュベートする</span></p>
+<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">結果</span></p>
+/29<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">翌日コンタミしてると考えられるビン</span><span lang="EN-US"><font face="Century">3</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">本培養に成功したビンが</span><span lang="EN-US"><font face="Century">3</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">本できた</span></p>
+(目的)<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">ただ念を入れて翌日同じ操作を行うことにした</span></p>
+大会提出用ベクターPsb1C3の培養とアルカリミニプレップ<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/30</font></span></p>
+(方法)<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">大会提出用ベクター</span><span lang="EN-US"><font face="Century">psb1c3</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の培養→トラフォ</span><span lang="EN-US"><font face="Century">(</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">二回目</span><span lang="EN-US"><font face="Century">)</font></span></p>
+/26に培養した提出用ベクターのコロニーの生えたプレートを用いた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">材料</span></p>
+↓LBプレート(amp+)に大腸菌をまき、37°C(API2-MALT1は30°C)で一晩培養した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">スクリュービン</span><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">本</span></p>
+↓プレートからシングルコロニーを分離した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">つまようじ</span><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">本</span></p>
+↓2 mlのLB培地で37°C(API2-MALT1は30°C)で一晩振とう培養した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/29</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">のコンピテント細胞を増やした</span><span lang="EN-US"><font face="Century">LB</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">培地</span></p>
+<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">LB</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">液体培地　</span><span lang="EN-US"><font face="Century">12ml</font></span></p>
+(結果)<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">クロラムフェ二コール　</span><span lang="EN-US"><font face="Century">12</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span></p>
+翌日コンタミしてると考えられるビン3本培養に成功したビンが3本できた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">方法</span></p>
+ただ念を入れて翌日同じ操作を行うことにした。<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓クリーンベンチにて昨日のプレートにマジックでコロニーに</span><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">つ印を入れる</span></p>
+<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓スクリュービン一本に液体培地</span><span lang="EN-US"><font face="Century">2ml</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">、クロラムフェ二コール</span><span lang="EN-US"><font face="Century">2</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">加えたものを</span><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">本作る</span></p>
+/30<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓火であぶったつまようじで先ほど印をつけたコロニーをつつきスクリュービンに入れるこれを</span><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">本分行う</span></p>
+(目的)<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓その後</span><span lang="EN-US"><font face="Century">37</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で一晩振とう培養する</span></p>
+大会提出用ベクターpsb1c3の培養→トラフォ(二回目)
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">結果</span></p>
+(方法)<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">本とも無事培養に成功し、コンタミも無かった</span></p>
+/26に培養した提出用ベクターのコロニーの生えたプレートを昨日同様用いた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
+↓LBプレート(amp+)に大腸菌をまき、37°C(API2-MALT1は30°C)で一晩培養した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/31</font></span></p>
+↓プレートからシングルコロニーを分離した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">培養した菌液のアルカリミニプレップ、フェノールクロロホルム処理、電気泳動、制限酵素処理</span></p>
+↓2 mlのLB培地で37°C(API2-MALT1は30°C)で一晩振とう培養した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">材料</span></p>
+<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
+(結果)<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">昨日の大腸菌が増えた菌液を取り出し</span><span lang="EN-US"><font face="Century">1.5ml</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">チューブに</span><span lang="EN-US"><font face="Century">1.4ml</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">程度入れる。その後</span><span lang="EN-US"><font face="Century">15000pm,5min,4</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で遠心</span></p>
+翌日6本培養したがコンタミすることなく無事に大腸菌を回収することができた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">上澄みを廃棄し</span><span lang="EN-US"><font face="Century">glucose solution(sol1)</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">を</span><span lang="EN-US"><font face="Century">100</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μｌ加えて</span><span lang="EN-US"><font face="Century">voltex</font></span></p>
+<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">沈殿物を溶かした後</span><span lang="EN-US"><font face="Century">0.2NNaCl1%SDS200</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μｌ加えてよく混ぜる</span></p>
+/31<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">その後氷上</span><span lang="EN-US"><font face="Century">5</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分</span></p>
+(目的)<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">氷冷した</span><span lang="EN-US"><font face="Century">3M</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の酢酸カリウムをいれて氷上五分</span></p>
+培養した菌液のアルカリミニプレップ、フェノールクロロホルム処理、電気泳動、制限酵素処理<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">15000rpm10min4</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で遠心</span></p>
+アルカリミニプレを最初に行った。<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">上澄みを新しいチューブに移す　入れて</span><span lang="EN-US"><font face="Century">voltex</font></span></p>
+<tr><td><table border=1 width="280px">
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">900</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の</span><span lang="EN-US"><font face="Century">2</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">‐プロパノールを加え混ぜて、二分待つ</span></p>
+<tr><td width="80px" align=center>Solution I</td><td width="200px"  align=right>50 mM グルコース (MW 180)</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">15000rpm,4</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃</span><span lang="EN-US"><font face="Century">,10</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分で遠心し、上澄みを捨てる</span></p>
+<tr><td>&nbsp;</td><td align=right>10 mM EDTA(pH 8.0)</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">１</span><span lang="EN-US"><font face="Century">ml</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の</span><span lang="EN-US"><font face="Century">70%</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">エタノールを加え</span><span lang="EN-US"><font face="Century">15000rpm,2</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分</span><span lang="EN-US"><font face="Century">,4</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で遠心する</span></p>
+<tr><td>&nbsp;</td><td align=right>25 mM Tris-HCl (pH 8.0)</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">沈殿物を乾かす</span></p>
+<tr><td align=center>Solution II</td><td align=right>0.2 N NaOH</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
+<tr><td>&nbsp;</td><td align=right>1% SDS</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">9/1<span style="mso-spacerun: yes">&nbsp;
+<tr><td align=center>Solution III</td><td align=right>3 M 酢酸カリウム</td></tr>
-</span></font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">ベクターのゲル抽出</span><span lang="EN-US"><font face="Century"> GFP</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">改良版の</span><span lang="EN-US"><font face="Century">PCR</font></span></p>
+<tr><td>&nbsp;</td><td align=right>1.8 M 酢酸</td></tr>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">材料</span></p>
+</table>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">1%</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">アガロースゲル</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">1kbp</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">マーカー</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">×</span><span lang="EN-US"><font face="Century">loading dye</font></span></p>
+↓LBプレート(amp+)に大腸菌をまき、37°C(API2-MALT1は30°C)で一晩培養した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">方法</span></p>
+↓プレートからシングルコロニーを分離した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓アガロース</span><span lang="EN-US"><font face="Century">0.2g</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">、</span><span lang="EN-US"><font face="Century">20</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">をまぜて</span><span lang="EN-US"><font face="Century">1%</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">アガロースゲルを作る</span></p>
+↓2 mlのLB培地で37°C(API2-MALT1は30°C)で一晩振とう培養した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓制限酵素処理したベクターを</span><span lang="EN-US"><font face="Century">50</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">と</span><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">×</span><span lang="EN-US"><font face="Century">loading dye</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">を</span><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">混ぜたものと</span><span lang="EN-US"><font face="Century">1kbp</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">のマーカーをゲルのコームに入れる</span></p>
+↓1.5 mlの培養液を1.5 mlチューブにうつし、15,000 rpm、4°Cで5分間遠心し、上清を捨てた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓</span><span lang="EN-US"><font face="Century">50V</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">、</span><span lang="EN-US"><font face="Century">60</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分電気泳動する</span></p>
+↓100 µlの氷冷したSolution Iを加え、ボルテックスした<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓ゲルを</span><span lang="EN-US"><font face="Century">EtBr</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">で</span><span lang="EN-US"><font face="Century">15</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分染色する</span></p>
+↓沈殿物を溶かした後、200 µlのSolution IIを加え、混ぜた、ボルテックスは使用しない<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓ゲルを紫外線に当てバンドのギリギリのところでカットする</span></p>
+↓氷上で5分間冷やした<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓それを</span><span lang="EN-US"><font face="Century">1.5ml</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">チューブに入れる</span></p>
+↓150 µlのSolution IIIを加え、混ぜた、ボルテックスは使用しない<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓重さを量りそれの三倍量の</span><span lang="EN-US"><font face="Century">Buffer QZ</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">を入れる</span></p>
+↓氷上で5分間冷やした<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓</span><span lang="EN-US"><font face="Century">50</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">度で</span><span lang="EN-US"><font face="Century">10</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分溶かす（時々</span><span lang="EN-US"><font face="Century">Voltex</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">）</span></p>
+↓15,000 rpm、4°Cで10分間遠心した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓ゲルと等量のイソプロパノールを加える</span></p>
+↓上清を新しいチューブにとり、沈殿物は廃棄した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓カラムに入れて</span><span lang="EN-US"><font face="Century">10000rpm</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">で一分遠心する</span></p>
+↓450 µlのイソプロパノールを加え、混ぜた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓</span><span lang="EN-US"><font face="Century">0.5ml</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の</span><span lang="EN-US"><font face="Century">Buffer QZ</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">をくわえて</span><span lang="EN-US"><font face="Century">10000rpm</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">で一分遠心</span></p>
+↓2分間室温で放置した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓</span><span lang="EN-US"><font face="Century">Buffer PE</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">を</span><span lang="EN-US"><font face="Century">1ml</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">くわえて</span><span lang="EN-US"><font face="Century">10000rpm</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">で一分遠心</span></p>
+↓15,000 rpm、室温で10分間遠心し、上清を捨てた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓再度</span><span lang="EN-US"><font face="Century">10000rpm</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">で遠心し水気をとる</span></p>
+↓150～200 µlの70%エタノールを加え、軽くボルテックスした<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓</span><span lang="EN-US"><font face="Century">MillQ</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">を</span><span lang="EN-US"><font face="Century">25</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">加えて</span><span lang="EN-US"><font face="Century">10000rpm</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">で一分遠心する</span></p>
+↓2分間室温で放置した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
+↓15,000 rpm、室温で10分間遠心し、上清を捨てた<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">PCR</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の</span></p>
+↓沈殿を10分～15分乾かした<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
+↓RNaseのはいったTEを20 µl加えて37°Cで培養した<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">9/2 GFP</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">のフェノクロ処理　電気泳動、制限酵素処理</span></p>
+<BR>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">フェノクロ処理</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">前回泳動した残りサンプルを合わせて</span><span lang="EN-US"><font face="Century">135</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">にする</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">milliQ65</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">加えて全量を</span><span lang="EN-US"><font face="Century">200</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">にする</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">フェノール</span><span lang="EN-US"><font face="Century">/</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">クロロホルムを等量入れる</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">しっかり</span><span lang="EN-US"><font face="Century">voltex</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">して、</span><span lang="EN-US"><font face="Century">15000rpm,5</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分</span><span lang="EN-US"><font face="Century">,25</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で遠心する</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">水層を別のエッペンに入れる（下の層がフェノール層）</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">次にクロロホルムのみで同様におこない、水層を別のエッペンに移す</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">1/10</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">等量の</span><span lang="EN-US"><font face="Century">3M</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">酢酸ナトリウムを加える</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">等量のイソプロパノールを加え、よく振る</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">12000rpm,20</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分</span><span lang="EN-US"><font face="Century">25</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で遠心する</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">沈殿をとらないように上澄みだけ除く</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">70%</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">エタノールを</span><span lang="EN-US"><font face="Century">200</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">加える</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">軽く混ぜた後</span><span lang="EN-US"><font face="Century">12000rpm,10</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">分</span><span lang="EN-US"><font face="Century">,25</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃で遠心</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">上澄みを除いた後乾燥させて長期保存なら</span><span lang="EN-US"><font face="Century">TE</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">、すぐに使うなら</span><span lang="EN-US"><font face="Century">milliQ50</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">に溶かす</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">制限酵素処理</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">先ほどの</span><span lang="EN-US"><font face="Century">DNA</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">溶液</span><span lang="EN-US"><font face="Century">50</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">と切りたい場所の制限酵素（今回は</span><span lang="EN-US"><font face="Century">Pst1,EcoR1</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">）を</span><span lang="EN-US"><font face="Century">1</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">、</span><span lang="EN-US"><font face="Century">Buffer</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">を使用する制限酵素にあわせたものを</span><span lang="EN-US"><font face="Century">6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">（今回は</span><span lang="EN-US"><font face="Century">10</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">×</span><span lang="EN-US"><font face="Century">H Buffer</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">）、</span><span lang="EN-US"><font face="Century">milliQ2</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">μ</span><span lang="EN-US"><font face="Century">l</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">を混ぜたものを</span><span lang="EN-US"><font face="Century">37</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">℃でインキュベートする</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">9/3 GFP</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">のゲル抽出</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">9/5 </font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">ベクターと</span><span lang="EN-US"><font face="Century">GFP</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">のライゲーションとトラフォ</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">9/6</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">二回目のベクターと</span><span lang="EN-US"><font face="Century">GFP</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">のライゲーション</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">9/7MLF</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の</span><span lang="EN-US"><font face="Century">PCR</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">と制限酵素処理とベクターの植菌</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">9/8MLF</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の完成、ライゲーションの相談とベクターのアルカリミニプレッツ、フェノクロ処理、制限酵素処理</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">9/9GFP</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">とベクターの濃度を上げる目的でのエタノール沈殿失敗</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">9/12</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">一回目大腸菌の植え付け</span><span lang="EN-US"><font face="Century"> GFP</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の</span><span lang="EN-US"><font face="Century">PCR</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">とフェノクロ処理と制限酵素処理</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">9/13</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">二回目大腸菌の植え付け　</span><span lang="EN-US"><font face="Century">GFP</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">のゲル電気泳動と抽出</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">9/14</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">大腸菌のベクター入りをアルカリミニプレッツ、フェノクロ処理、制限酵素処理</span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
-<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">9/15 </font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">大腸菌のベクターを電気泳動、ゲル抽出、二回目</span><span lang="EN-US"><font face="Century">GFP</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">＋ベクターのライゲーション</span></p>
-<p>&nbsp;</p>
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