https://eln.excellgen.com/index.php?action=history&feed=atom&title=1026-1028-pET28b_lambda_exo_beta_gam 1026-1028-pET28b lambda exo beta gam - Revision history 2026-10-18T02:24:15Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1026-1028-pET28b_lambda_exo_beta_gam&diff=7804&oldid=prev Paul.xu at 21:25, 9 January 2013 2013-01-09T21:25:05Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1026~1028, pET28b lambda alpha (exo), beta, gama&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of lambda RED (exo and related proteins)&lt;br&gt; <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2012-3-25- <br /> <br /> &#039;&#039;&#039;Notebook:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:1026&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; yingying <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === cloning ===<br /> <br /> date: 3/25/2012&lt;br&gt; <br /> <br /> PCR using phage DNA &lt;br&gt; <br /> <br /> {| cellspacing=&quot;1&quot;<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> [[Image:Lambda phage PCR 3 25 2012.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> 3/28/2012 <br /> <br /> Miniprep DigestionNde1+ Xho1&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:Lambda miniprep digestion 3 28 2012.jpg]] <br /> <br /> [[Image:Lambda miniprep digestion2 3 28 2012.jpg]] <br /> <br /> sequencing: L1-3 (no priming), L2-1 (bad quality), L3-5 (too many mutations) <br /> <br /> L1 is lambda alpha, 650bp <br /> <br /> L2 is lambda gama, 400 bp <br /> <br /> L3 is lamda beta, 700bp <br /> <br /> &lt;br&gt; <br /> <br /> 3/29/2012, colony PCR screening <br /> <br /> [http://www.excellgen.net/eln/images/5/58/1026_1530_1533_1565pcr.JPG 1026_1530_1533_1565pcr.JPG]‎ [[Image:Lambda colony PCR screening1 3 29 2012.jpg]] <br /> <br /> [[Image:Lambda PCR screening2 3 29 2012.jpg]] <br /> <br /> 3/30/2012Make new miniprep, digestion with Nde1+ Xho1&lt;br&gt; <br /> <br /> [http://www.excellgen.net/eln/images/5/58/1026_1530_1533_1565pcr.JPG 1026_1530_1533_1565pcr.JPG]‎ [http://www.excellgen.net/eln/images/5/58/1026_1530_1533_1565pcr.JPG 1026_1530_1533_1565pcr.JPG]‎ [[Image:Lambda exo bet gam digestion Nde1 Xho1 2012 3 30.JPG]] <br /> <br /> exo:&amp;nbsp; left 4&lt;br&gt; <br /> <br /> bet: middle 4 <br /> <br /> gam: right 4 <br /> <br /> send sequencing, left 1, middle 2, right 2 <br /> <br /> &lt;br&gt; <br /> <br /> 4/10/2012 select 4 clones to mini-prep, digest &lt;br&gt; <br /> <br /> [[Image:1012 1078 1026 1027 digest 4.10.2012.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Cloning of 1026 pET28b-LambdaExo&#039;&#039;&#039; ==<br /> <br /> === cloning ===<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1026 pET28b-LambdaExo&lt;br&gt; <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-1-3- <br /> <br /> &#039;&#039;&#039;Notebook:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:1026&#039;&#039;&#039; <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 phage DNA (25cycles)&lt;br&gt; <br /> <br /> [[Image:1026 1530 1533 1565pcr.JPG]]&lt;br&gt; <br /> <br /> [[Image:1026pcr v12digest.JPG]]&lt;br&gt; <br /> <br /> === colony screening ===<br /> <br /> &lt;br&gt; Miniprep Digestion Nde1+ Xho1&lt;br&gt; <br /> <br /> [[Image:1026_1570_1565_1501_1533DIGEST.JPG]]<br /> <br /> &lt;br&gt; <br /> <br /> [[Category:Cloning]]</div> Paul.xu