Team:KIT-Kyoto/Protocol
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+ | <table width="700px" align=center> | ||
+ | <tr><td width="340px"> | ||
+ | <span id="lig">'''ライゲーション'''</span></td><td width="20px"> </td><td width="340px"> | ||
+ | <span id="ligeng">'''DNA ligation'''</span></td></tr> | ||
+ | <tr><td>[https://www.roche-applied-science.com/servlet/RCProductDisplay?storeId=10202&catalogId=10202&langId=-1&countryId=us&forCountryId=us&productId=3.6.14.2.2.3 Rapid DNA Ligation Kit]を使用する</td><td> </td><td>We use [https://www.roche-applied-science.com/servlet/RCProductDisplay?storeId=10202&catalogId=10202&langId=- | ||
+ | 1&countryId=us&forCountryId=us&productId=3.6.14.2.2.3 Rapid DNA Ligation Kit].</td></tr> | ||
+ | <tr><td>↓ 下記の組成に従って試薬を混ぜる</td><td> </td><td>>↓ Mix the reagent according to the following components.</td></tr> | ||
+ | <tr><td><table border=1 width="300px"><tr><td align=center width="200px">5 x DNA dilution buffer</td><td align=right width="100px">2 μl</td></tr> | ||
+ | <tr><td align=center>ベクター(カット・CHIP処理後)</td><td align=right>0.25-1 μl</td></tr> | ||
+ | <tr><td align=center>インサート DNA</td><td align=right>0.5-6.5 μl</td></tr> | ||
+ | <tr><td align=center>ddH2O</td><td align=right>全量が10 μlになるように調整する</td></tr></table></td><td> </td><td><table border=1 wight="300px"><tr><td wight="200px" align=center>5x DNA dilution buffer</td><td wight="100px" align=right>2 μl</td></tr> | ||
+ | <tr><td align=center>Vector(cut and CHIP treated)</td><td align=right>0.25-1 μl</td></tr> | ||
+ | <tr><td align=center>Insert DNA</td><td align=right>0.5-6.5 μl</td></tr> | ||
+ | <tr><td align=center>ddH2O</td><td align=right>To final volume of 10 μl</td></tr></table></td></tr> | ||
+ | <tr><td>↓ よく混ぜる</td><td> </td><td>↓ Mix well.</td></tr> | ||
+ | <tr><td>↓ 10 μlの2 x rapid ligation bufferと1 μlのligaseを加える</td><td> </td><td>↓ Add 10 μl of | ||
+ | 2 x rapid ligation buffer and 1 μl ligase.</td></tr> | ||
+ | <tr><td>↓ よく混ぜる</td><td> </td><td>↓ Mix well.</td></tr> | ||
+ | <tr><td>↓ 室温で5分間放置する</td><td> </td><td>↓ Hold at room temperature for 5 minutes.</td></tr> | ||
+ | <tr><td>↓ トランスフォーメーションに使用するときは、100 μlのコンピテント細胞に対して10 μl使用する</td><td> </td><td>↓Then use 10 μl of the ligation reaction to transform 100 μl of chemically competent cells.</td></tr> | ||
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+ | </table> | ||
</td></tr> | </td></tr> | ||
+ | <tr><td> | ||
+ | <div align="right"><span style="font-size:10pt;">[[#Contents|>>back to Contents]]</span></div> | ||
- | + | </td></tr> | |
Revision as of 05:18, 24 October 2010
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About protocol
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