Team:KIT-Kyoto/nakagawa

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<p><strong>LBプレートの作成法<br>
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<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/22</font></span></p>
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<br>
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<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>
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</strong>↓下記の組成に従って試薬を混ぜる
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<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
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<table style="WIDTH: 261px; HEIGHT: 102px" border="1" cellspacing="0" cellpadding="0" width="261">
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<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>
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  <tr>
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<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>
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    <td> NaCl</td>
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<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>
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    <td>
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<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>
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      <p align="left">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 2.5g </p></td>
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<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>
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  </tr>
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<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>
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  <tr>
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<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
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    <td> Bacto Agar</td>
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<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/23</font></span></p>
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    <td> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;7.5g</td>
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<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>
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  </tr>
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<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>
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  <tr>
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<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>
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    <td> Bacto Yeast Extract</td>
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<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>
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    <td> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 2.5g</td>
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<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>
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  </tr>
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<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
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  <tr>
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<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>
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    <td> Bacto Trypton</td>
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<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
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    <td> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 5.0g</td>
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<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/24</font></span></p>
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  </tr>
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<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>
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  <tr>
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<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
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    <td> H<font size="1">2</font>O</td>
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<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>
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    <td> &nbsp;&nbsp;&nbsp;  
+
<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>
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500ml</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">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>
-
  </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">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>
+
<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>
-
</p>↓120℃で20分間オートクレーブする <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>
-
<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>
-
 
+
<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>
-
↓ある程度冷めたらクリーンベンチでシャーレを並べ、<br>
+
<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>
-
デカントで入れる。<LBchl<sup>+</sup>  
+
<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>
-
の時50mg/<font size="2">μ</font>lのchlを0.5ml<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">沈殿物を乾かす</span></p>
-
加える><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">電気泳動</span></p>
-
 
+
<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><o:p><font face="Century"></font></o:P></span></p>
-
<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>
-
↓シャーレの気泡をバーナーの火でなくす<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">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>
-
 
+
<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>
-
<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">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>
-
↓固まったら37℃でインキュベートする<br>
+
<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>
-
 
+
<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>
-
<br>
+
<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>
-
↓冷蔵保管する
+
<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>
-
<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 lang="EN-US"><font face="Century">8/25</font></span></p>
-
 
+
<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>
 +
<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">PCR</font></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">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>
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<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>
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<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>
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<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>
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<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>
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<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"><font face="Century">8/26</font></span></p>
 +
<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>
 +
<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/28</font></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">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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/29</font></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">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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<p style="MARGIN: 0mm 0mm 0pt" class="MsoNormal"><span lang="EN-US"><font face="Century">8/30</font></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">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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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">8/31</font></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">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>
 +
<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>
 +
<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">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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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">9/1<span style="mso-spacerun: yes">&nbsp;  
 +
</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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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>
 +
<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">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>
 +
<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">9/2 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>
 +
<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>
</body>
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Revision as of 13:12, 18 September 2011

8/22

トランスフォーメーション

↓氷上でコンピテント細胞(DH5 Alpha 大腸菌体)解凍

↓前もって冷やしておいた1.5mlチューブに100μlのコンピテント細胞を分注する

DNAをチューブに1~5μl加えて氷上で30分正確に冷やす

42℃で45秒熱ショックを与える

0.1mlLBプレート(+amp)にまく

37度で一晩培養

8/23

PCRプライマー設計 プレカルチャー

大腸菌のベクターにはEXSPの制限酵素で切れる場所がありEX DNA SPとなる

今回はまずE2~3X2~3S3P何塩基でもつけてよい。そしてGFPにつなげる上で15塩基必要なのでその条件で設計する。

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。

そしてプライマー作成サイトでは温度が65度を超えないように注意する

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6本分し、2mlずつLBの液体培地に入れて37℃で一晩培養(+amp

8/24

アルカリミニプレッツ(プラスミド回収)

昨日の大腸菌が増えた菌液を取り出し1.5mlチューブに1.4ml程度入れる。その後15000pm,5min,4℃で遠心

上澄みを廃棄しglucose solution(sol1)100μl加えてvoltex

沈殿物を溶かした後0.2NNaCl1%SDS200μl加えてよく混ぜる

その後氷上5

氷冷した3Mの酢酸カリウムをいれて氷上五分

15000rpm10min4℃で遠心

上澄みを新しいチューブに移す 入れてvoltex

900μl2‐プロパノールを加え混ぜて、二分待つ

15000rpm,4,10分で遠心し、上澄みを捨てる

ml70%エタノールを加え15000rpm,2,4℃で遠心する

沈殿物を乾かす

電気泳動

6本の乾燥DNAにwilliQ0.3ml DNA5μlEcoR10.5μlPst10.5μl10×Buffer 0.7μl

加え3730分インキュベート

%agaゲル20μlより0.2gagaを使いそこにTMN20ml加えてゲルとする

電子レンジで沸騰させて粒がなくなった後約20分待つ

1kbaseのマーカー5μlDNAのマーカー(×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ぐ

電気泳動 100V 30

EtBr染色(手袋をつけて) 10

撮影

8/25

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

PCR

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にする

milliQ65μl加えて全量を200μlにする

フェノール/クロロホルムを等量入れる

しっかりvoltexして、15000rpm,5,25℃で遠心する

水層を別のエッペンに入れる(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す

1/10等量の3M酢酸ナトリウムを加える

等量のイソプロパノールを加え、よく振る

12000rpm,2025℃で遠心する

沈殿をとらないように上澄みだけ除く

70%エタノールを200μl加える

軽く混ぜた後12000rpm,10,25℃で遠心

上澄みを除いた後乾燥させて長期保存ならTE、すぐに使うならmilliQ50μlに溶かす

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1μlBufferを使用する制限酵素にあわせたものを6μl(今回は10×H Buffer)、milliQ2μlを混ぜたものを37℃でインキュベートする

8/26

GFPゲル抽出

8/28

大腸菌のコンピテント細胞に提出用ベクターPsb1c3を入れてインキュベート

材料

大腸菌のコンピテント細胞 100μl

クロラムフェ二コール   10μl

LBプレート

大会提出用ベクターpSB1C3 1μl

方法

↓提出用ベクターpSB1C3の培養

↓クリーンベンチにて大腸菌のコンピテント細胞を1.5mlのチューブに取る

↓そこにクロラムフェ二コールを10μlpSB1C31μl加える

↓それをLBプレートにまき、ガラス棒でのばす

37℃で一晩インキュベートする

結果

小さいながらもコロニーを確認することが出来た

8/29

大会提出用ベクターpsb1c3の培養→トラフォ(一回目)

材料

スクリュービン6

つまようじ6

8/29のコンピテント細胞を増やしたLB培地

LB液体培地 12ml

クロラムフェ二コール 12μl

方法

↓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6つ印を入れる

↓スクリュービン一本に液体培地2ml、クロラムフェ二コール2μl加えたものを6本作る

↓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6本分行う

↓その後37℃で一晩インキュベートする

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

大会提出用ベクターpsb1c3の培養→トラフォ(二回目)

材料

スクリュービン6

つまようじ6

8/29のコンピテント細胞を増やしたLB培地

LB液体培地 12ml

クロラムフェ二コール 12μl

方法

↓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6つ印を入れる

↓スクリュービン一本に液体培地2ml、クロラムフェ二コール2μl加えたものを6本作る

↓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6本分行う

↓その後37℃で一晩振とう培養する

結果

6本とも無事培養に成功し、コンタミも無かった

8/31

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

材料

昨日の大腸菌が増えた菌液を取り出し1.5mlチューブに1.4ml程度入れる。その後15000pm,5min,4℃で遠心

上澄みを廃棄しglucose solution(sol1)100μl加えてvoltex

沈殿物を溶かした後0.2NNaCl1%SDS200μl加えてよく混ぜる

その後氷上5

氷冷した3Mの酢酸カリウムをいれて氷上五分

15000rpm10min4℃で遠心

上澄みを新しいチューブに移す 入れてvoltex

900μl2‐プロパノールを加え混ぜて、二分待つ

15000rpm,4,10分で遠心し、上澄みを捨てる

ml70%エタノールを加え15000rpm,2,4℃で遠心する

沈殿物を乾かす

9/1  ベクターのゲル抽出 GFP改良版のPCR

材料

1%アガロースゲル

1kbpマーカー

6×loading dye

方法

↓アガロース0.2g20μlをまぜて1%アガロースゲルを作る

↓制限酵素処理したベクターを50μl6×loading dye6μl混ぜたものと1kbpのマーカーをゲルのコームに入れる

50V60分電気泳動する

↓ゲルをEtBr15分染色する

↓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

↓それを1.5mlチューブに入れる

↓重さを量りそれの三倍量のBuffer QZを入れる

50度で10分溶かす(時々Voltex

↓ゲルと等量のイソプロパノールを加える

↓カラムに入れて10000rpmで一分遠心する

0.5mlBuffer QZをくわえて10000rpmで一分遠心

Buffer PE1mlくわえて10000rpmで一分遠心

↓再度10000rpmで遠心し水気をとる

MillQ25μl加えて10000rpmで一分遠心する

PCR

9/2 GFPのフェノクロ処理 電気泳動、制限酵素処理

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にする

milliQ65μl加えて全量を200μlにする

フェノール/クロロホルムを等量入れる

しっかりvoltexして、15000rpm,5,25℃で遠心する

水層を別のエッペンに入れる(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す

1/10等量の3M酢酸ナトリウムを加える

等量のイソプロパノールを加え、よく振る

12000rpm,2025℃で遠心する

沈殿をとらないように上澄みだけ除く

70%エタノールを200μl加える

軽く混ぜた後12000rpm,10,25℃で遠心

上澄みを除いた後乾燥させて長期保存ならTE、すぐに使うならmilliQ50μlに溶かす

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1μlBufferを使用する制限酵素にあわせたものを6μl(今回は10×H Buffer)、milliQ2μlを混ぜたものを37℃でインキュベートする

9/3 GFPのゲル抽出

9/5 ベクターとGFPのライゲーションとトラフォ

9/6二回目のベクターとGFPのライゲーション

9/7MLFPCRと制限酵素処理とベクターの植菌

9/8MLF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

9/9GFP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

9/12一回目大腸菌の植え付け GFPPCRとフェノクロ処理と制限酵素処理

9/13二回目大腸菌の植え付け GFPのゲル電気泳動と抽出

9/14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

9/15 大腸菌のベクターを電気泳動、ゲル抽出、二回目GFP+ベクターのライゲーション