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Team:KIT-Kyoto/yokoigawa
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| - | < | + | <p style="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><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">月</span><span lang="EN-US"><font face="Century">)</font></span></p> |
| - | + | <p style="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">武田、横井川</span></p> | |
| - | + | <p style="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">【目的】</span></p> | |
| - | + | <p style="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><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">産物と</span><span lang="EN-US"><font face="Century">pUAST-flag vector</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">の制限酵素処理（一回目：</span><span lang="EN-US"><font face="Century">Xho 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">）</span></p> | |
| - | + | <p style="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">【実験操作】</span></p> | |
| - | + | <p style="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">～フェノールクロロホルム処理・エタノール沈殿～</span></p> | |
| - | + | <p style="TEXT-ALIGN: left; TEXT-INDENT: -10.5pt; MARGIN: 0mm 0mm 0pt 10.5pt; mso-char-indent-count: -1.0" class="MsoNormal" align="left"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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">DAIP2</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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">産物を</span><span lang="EN-US"><font face="Century">400</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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">まで</span><span lang="EN-US"><font face="Century">milliQ</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">で希釈し、フェノール・クロロホルムを</span><span lang="EN-US"><font face="Century">400</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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">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</span><span lang="EN-US"><font face="Century">10000rpm3min4</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓水層のみを新しいチューブに移した。</span></p> | |
| - | + | <p style="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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加えて、</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</span><span lang="EN-US"><font face="Century">10000rpm3min4</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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="TEXT-ALIGN: left; TEXT-INDENT: -10.5pt; MARGIN: 0mm 0mm 0pt 10.5pt; mso-char-indent-count: -1.0" class="MsoNormal" align="left"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓水層のみを新しいチューブに移し</span><span lang="EN-US"><font face="Century">3M CH<sub>3</sub>COONa(PH 5.27)</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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">1/10</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">等量加え、</span><span lang="EN-US"><font face="Century">isopropanol</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">を等量加えた。</span></p> | |
| - | + | <p style="TEXT-ALIGN: left; TEXT-INDENT: -10.5pt; MARGIN: 0mm 0mm 0pt 10.5pt; mso-char-indent-count: -1.0" class="MsoNormal" align="left"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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">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">分静置後、</span><span lang="EN-US"><font face="Century">14000rpm 10min 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="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓遠心後、上清を捨て、</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">％</span><span lang="EN-US"><font face="Century">ethanol</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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">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、</span><span lang="EN-US"><font face="Century">15000rpm 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="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">↓遠心後、上清を捨て乾燥させる。乾燥後</span><span lang="EN-US"><font face="Century">milliQ44</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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">を加えて</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、</span><span lang="EN-US"><font face="Century">milliQ</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">中に</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">を溶解させる。</span></p> | |
| - | + | <p style="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">フェノールクロロホルム処理後下記の反応液を調整した</span></p> | |
| - | + | <p style="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt 21pt; mso-para-margin-left: 0gd" class="MsoListParagraph" align="left"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin">反応液調整（</span><span lang="EN-US"><font face="Century">cDNA</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt 21pt; mso-para-margin-left: 0gd" class="MsoListParagraph" align="left"><span lang="EN-US"><font face="Century"><span style="mso-spacerun: yes"> </span>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">産物</span><span lang="EN-US"><font face="Century"> in milliQ<span style="mso-spacerun: yes"> | |
| - | + | </span>44</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt 21pt; mso-para-margin-left: 0gd" class="MsoListParagraph" align="left"><span lang="EN-US"><font face="Century"><span style="mso-spacerun: yes"> </span>10x 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">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></p> | |
| - | + | <p style="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt 21pt; mso-para-margin-left: 0gd" class="MsoListParagraph" align="left"><span lang="EN-US"><font face="Century"><span style="mso-spacerun: yes"> </span>Xho 1<span style="mso-spacerun: yes"> | |
| - | </ | + | </span>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="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt 21pt; mso-para-margin-left: 0gd" class="MsoListParagraph" align="left"><span lang="EN-US"><?xml:namespace prefix ="" o ns | ||
| + | ="" "urn:schemas-microsoft-com:office:office" /><o:p><font face="Century"> </font></o:p></span></p> | ||
| + | <p style="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin"> 反応液調整（</span><span lang="EN-US"><font face="Century">vector</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><span lang="EN-US"><font face="Century"><span style="mso-spacerun: yes"> </span>pUAST-flag | ||
| + | vector(500ng/</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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)<span style="mso-spacerun: yes"> | ||
| + | </span>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="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><span lang="EN-US"><font face="Century"><span style="mso-spacerun: yes"> </span>milliQ<span style="mso-spacerun: yes"> | ||
| + | </span>34</font></span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; 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="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><span lang="EN-US"><font face="Century"><span style="mso-spacerun: yes"> </span>10x H | ||
| + | Buffer<span style="mso-spacerun: yes"> | ||
| + | </span>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></p> | ||
| + | <p style="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><span lang="EN-US"><font face="Century"><span style="mso-spacerun: yes"> </span>Xho 1<span style="mso-spacerun: yes"> | ||
| + | </span>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="TEXT-ALIGN: left; MARGIN: 0mm 0mm 0pt" class="MsoNormal" align="left"><span lang="EN-US"><o:p><font face="Century"> </font></o:p></span></p> | ||
| + | <p><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; FONT-SIZE: 10.5pt; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin; mso-bidi-font-size: 11.0pt; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: JA; mso-bidi-language: AR-SA">↓反応液調整後、</span><span style="FONT-FAMILY: 'Century','serif'; FONT-SIZE: 10.5pt; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-theme-font: minor-latin; mso-bidi-font-size: 11.0pt; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: JA; mso-bidi-language: AR-SA" lang="EN-US">37</span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; FONT-SIZE: 10.5pt; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin; mso-bidi-font-size: 11.0pt; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: JA; mso-bidi-language: AR-SA">℃で</span><span style="FONT-FAMILY: 'Century','serif'; FONT-SIZE: 10.5pt; mso-ascii-theme-font: minor-latin; mso-fareast-font-family: 'ＭＳ 明朝'; mso-fareast-theme-font: minor-fareast; mso-hansi-theme-font: minor-latin; mso-bidi-font-size: 11.0pt; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: JA; mso-bidi-language: AR-SA" lang="EN-US">20</span><span style="FONT-FAMILY: 'ＭＳ 明朝','serif'; FONT-SIZE: 10.5pt; mso-ascii-font-family: Century; mso-ascii-theme-font: minor-latin; mso-fareast-theme-font: minor-fareast; mso-hansi-font-family: Century; mso-hansi-theme-font: minor-latin; mso-bidi-font-size: 11.0pt; mso-bidi-font-family: 'Times New Roman'; mso-bidi-theme-font: minor-bidi; mso-ansi-language: EN-US; mso-fareast-language: JA; mso-bidi-language: AR-SA">時間静置</span></p> | ||
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Revision as of 09:36, 18 September 2011
8/29(月)
武田、横井川
【目的】
PCR産物とpUAST-flag vectorの制限酵素処理（一回目：Xho 1）
【実験操作】
～フェノールクロロホルム処理・エタノール沈殿～
↓DAIP2のPCR産物を400μlまでmilliQで希釈し、フェノール・クロロホルムを400μl加えてvoltex し10000rpm3min4℃で遠心。
↓水層のみを新しいチューブに移した。
↓クロロホルムを等量加えて、voltexし10000rpm3min4℃で遠心。
↓水層のみを新しいチューブに移し3M CH3COONa(PH 5.27)を1/10等量加え、isopropanolを等量加えた。
↓3分静置後、14000rpm 10min 4℃で遠心。
↓遠心後、上清を捨て、70％ethanolを200μl加え、15000rpm 5min 4℃で遠心。
↓遠心後、上清を捨て乾燥させる。乾燥後milliQ44μlを加えてvoltexし、milliQ中にDNAを溶解させる。
～制限酵素処理～
フェノールクロロホルム処理後下記の反応液を調整した
反応液調整（cDNA）
PCR産物 in milliQ 44μl
10x H Buffer 5μl
Xho 1 1μl
反応液調整（vector）
pUAST-flag vector(500ng/μl) 10μl
milliQ 34μl
10x H Buffer 5μl
Xho 1 1μl
↓反応液調整後、37℃で20時間静置
