https://eln.excellgen.com/index.php?action=history&feed=atom&title=1127_pET30b-T4RNA-Lig2K227Q 1127 pET30b-T4RNA-Lig2K227Q - Revision history 2026-10-18T02:19:58Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1127_pET30b-T4RNA-Lig2K227Q&diff=8244&oldid=prev Paul.xu at 16:43, 15 February 2013 2013-02-15T16:43:00Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1127 pET30b-T4RNA-Lig2K227Q&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1127 pET30b-T4RNA-Lig2K227Q <br /> <br /> &#039;&#039;&#039;Date: 1, 2013&lt;br&gt; &#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Notebook:&amp;nbsp;&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID: &#039;&#039;&#039;1127 <br /> <br /> &#039;&#039;&#039;Experimenters: &#039;&#039;&#039; shanyi <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === Cloning ===<br /> <br /> PCR using T4 phage DNA <br /> <br /> 1: T4RNA-lig-Nde1-F + T4RNAlig-K227Q-R1 using T4 phage DNA as template, not more than 20 cycles <br /> <br /> 2: T4RNA-lig-Nde1-F + T4RNAlig-K227Q-R2 using PCR product from PCR reaction 1 as template&lt;br&gt; <br /> <br /> [[Image:1127pcrfirst.JPG]]&lt;br&gt; <br /> <br /> second round pcr&lt;br&gt; <br /> <br /> [[Image:1045 1049 1123 1127PCR.JPG]]&lt;br&gt; <br /> <br /> Clone into pET30b Nde1 + Xho1&lt;br&gt; <br /> <br /> [[Image:1514 1123pcr v16 1127 v12 1530digest.JPG]] <br /> <br /> ligate and transform, take kan plate,should use amp plate <br /> <br /> [[Image:1097 1127 1127 1045pcr.JPG.JPG]] <br /> <br /> [[Image:1097pcr 1279 1127 v15 1530pcr v12digest.JPG]] <br /> <br /> === &lt;u&gt;&lt;/u&gt;Colony Screening&lt;br&gt; ===<br /> <br /> ----<br /> <br /> [[Image:1097 1127 1530 1279 1533 1568 1590digest.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Cloning of 1127 pET30b-T4RNA-Lig2K227Q&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1127 pET30b-T4RNA-Lig2K227Q <br /> <br /> &#039;&#039;&#039;Date: 2, 2013&lt;br&gt; &#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Notebook:&amp;nbsp;&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID: &#039;&#039;&#039;1127 <br /> <br /> &#039;&#039;&#039;Experimenters: &#039;&#039;&#039; shanyi <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === Cloning ===<br /> <br /> PCR <br /> <br /> 1: T4RNA-lig-Nde1-F + T4RNAlig-K227Q-R1 using 1123 as template, not more than 20 cycles <br /> <br /> [[Image:1127first PCR.JPG]] <br /> <br /> 2: T4RNA-lig-Nde1-F + T4RNAlig-K227Q-R2 using PCR product from PCR reaction 1 as template&lt;br&gt; <br /> <br /> [[Image:11272nd round pcr.JPG]]&lt;br&gt; <br /> <br /> Clone into pET30b Nde1 + Xho1&lt;br&gt; <br /> <br /> [[Image:Digest for1127.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> ligate and transform, <br /> <br /> &lt;br&gt; <br /> <br /> === &lt;u&gt;&lt;/u&gt;Colony Screening&lt;br&gt; ===<br /> <br /> ----<br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &lt;br&gt; <br /> <br /> [[Category:Cloning]]</div> Paul.xu