https://eln.excellgen.com/index.php?action=history&feed=atom&title=1454_pEC 2026-10-18T06:19:44Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1454_pEC&diff=7093&oldid=prev 2012-11-16T20:05:28Z

<p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1454 pEC&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; pEC <br /> <br /> &#039;&#039;&#039;Date:&amp;nbsp;2012-11-8&#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:&amp;nbsp;1454&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; Shanyi <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039;&lt;br&gt; ==<br /> <br /> 11/7/2012, Digestion <br /> <br /> [[Image:1.2 1.6 2.1 digest.JPG]] <br /> <br /> Clone 1.2 is pCR2.1 EcoR1 digest and religate. <br /> <br /> &lt;br&gt; [[Image:16 Nco1 Alwn1 11 8 2012.jpg]][[Image:16 Alwn1 Xho1 11 8 2012.jpg]] <br /> <br /> &lt;br&gt; <br /> <br /> 1.6 clone 1 and 3 are not good, they are two copies of Amp-(or Amp-Kan) ligated from Alwn1-DraIII fragment from pCR2.1 <br /> <br /> 2.1 clone #4 is pRSetA. <br /> <br /> 2.1 Clone # 5 and 6 are from pRSET-A <br /> <br /> &lt;br&gt; <br /> <br /> 11-05-2012&lt;br&gt; <br /> <br /> subcloning&lt;br&gt; <br /> <br /> [[Image:V16 pCR2.1AND 1293P V15 414 1449 1445P 1449 11 05 2012.JPG]]&lt;br&gt; <br /> <br /> Digest 1457 pCR2.1 with DraIII + AlwN1---vector, cut out 1296 bp fragment&lt;br&gt;digest pET28b-amp with DraIII + AlwN1---insert,&lt;br&gt;ligate and transform, plate on LB/amp &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> Colony Screening&lt;br&gt; <br /> <br /> [[Image:1454.JPG]]&lt;br&gt; <br /> <br /> digest with \DraIII + AlwN1&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:1352 1298 1328 1338 1454fdigest11 14 2012.JPG]]&lt;br&gt; <br /> <br /> first 1454(from first step) digest with HindIII + EcoR1&lt;br&gt; <br /> <br /> the second 1454 (from first step) digest with Pst1&lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2012- <br /> <br /> Make miniprep and digest with DraIII + Nco1. <br /> <br /> [[Image:1312 1339 1454digest11 16 2012.JPG]] <br /> <br /> [[Image:1454digest11_16_2012.JPG]]<br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> &lt;br&gt; <br /> <br /> [[Category:Cloning]]</div>

Paul.xu