https://eln.excellgen.com/index.php?action=history&feed=atom&title=1530_pET28b-Klenow3%275exo%E2%80%93 1530 pET28b-Klenow3'5exo– - Revision history 2026-10-18T05:03:47Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1530_pET28b-Klenow3%275exo%E2%80%93&diff=8073&oldid=prev Paul.xu at 19:47, 30 January 2013 2013-01-30T19:47:33Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of pET28b-Klenow(3&#039;-&amp;gt;5&#039;_exo–)&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of pET28b-Klenow(3&#039;-&amp;gt;5&#039;_exo–) <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;1530 <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:two Fragments,then Join two fragment by PCR(25cycles) <br /> <br /> [[Image:1026 1530 1533 1565pcr.JPG]]&lt;br&gt; <br /> <br /> [[Image:1530 1533pcr 1 10 2013.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 /> digest with Nde1 + Sal1, clone into pET28b <br /> <br /> [[Image:1097 1127 1530 1279 1533 1568 1590digest.JPG]] <br /> <br /> [[Image:1049 1122 1530DIGEST.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Cloning of pET28b-Klenow(3&#039;-&amp;gt;5&#039;_exo–)&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of pET28b-Klenow(3&#039;-&amp;gt;5&#039;_exo–) <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;1530 <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:use 1117 as template,two Fragments,then Join two fragment by PCR(25cycles) <br /> <br /> [[Image:1530PCR 1 23 2013-2.JPG]]&lt;br&gt; <br /> <br /> [[Image:1530PCR 1 24 2013.JPG]]&lt;br&gt; <br /> <br /> cut out the first band(from up to down)&lt;br&gt; <br /> <br /> digest with Nde1 + Sal1,clone into pET28b &lt;br&gt; <br /> <br /> [[Image:DIGEST FOR 1601 1528 1530 1 25 2013.JPG]]&lt;br&gt; <br /> <br /> cut out the first band(from up to down)and the digested vector&lt;br&gt; <br /> <br /> === &lt;u&gt;&lt;/u&gt;Colony Screening&lt;br&gt; ===<br /> <br /> digest with Nde1 + Sal1,<br /> <br /> [[Image:549 1530 1501digest.JPG]]&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