https://eln.excellgen.com/index.php?action=history&feed=atom&title=1619-pET28b-T4DNAPol 1619-pET28b-T4DNAPol - Revision history 2026-10-18T03:06:00Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1619-pET28b-T4DNAPol&diff=9611&oldid=prev Paul.xu at 15:30, 3 July 2013 2013-07-03T15:30:51Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1619-pET28b-T4DNAPol &#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1619-pET28b-T4DNAPol <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-3-25---2013-3-29 <br /> <br /> &#039;&#039;&#039;Notebook:0&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:1619&#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 /> [[Image:1619 PCR 3-26-2013.JPG]] <br /> <br /> [[Image:1619+1704v+1710 clone 3-27-2013.JPG]] <br /> <br /> lane1: PCR product <br /> <br /> lane2: pET28b vector <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2013-3-29 <br /> <br /> Make miniprep and digest with Nhe1 + Not1 <br /> <br /> [[Image:1619 digest detect 3-29-2013.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 /> == &#039;&#039;&#039;Cloning of 1619-pET28b-T4DNAPol&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039;&amp;nbsp; Fix of Cloning of 1619-pET28b-T4DNAPol <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-4-29--2013-5-1 <br /> <br /> &#039;&#039;&#039;Notebook:0&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:1619&#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 /> [[Image:1619 pcr1+2 4-11-2013.JPG]]&lt;br&gt; <br /> <br /> template:1619-4 above <br /> <br /> pcr1primer: 1-1 to 3-8 <br /> <br /> pcr2 primer: 3-9 to 3-24 <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:1619 PCR 4-19-2013.JPG]]&lt;br&gt; <br /> <br /> template: pcr1+pcr2 <br /> <br /> primer: 1-1 to 3-24 <br /> <br /> clone PCR product to pET28b <br /> <br /> == Colony Screening ==<br /> <br /> Date: 2013-5-1 <br /> <br /> Make miniprep and digest Nhe1 + Not1 &lt;br&gt; <br /> <br /> [[Image:1619 digest detect 5-1-2013.JPG]] <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> *&amp;lt;br&amp;gt;<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;fix of Cloning of 1619-pET28b-T4DNAPol&#039;&#039;&#039;&lt;br&gt; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039;&amp;nbsp; Fix of Cloning of 1619-pET28b-T4DNAPol <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-5-6---2013-5-23 <br /> <br /> &#039;&#039;&#039;Notebook:0&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:1619&#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 /> [[Image:1619 pcr1+pcr2 5-6-2013.JPG]]&lt;br&gt; <br /> <br /> template:1619-1 (above2013/5/1)&lt;br&gt; <br /> <br /> pcr1 primer: 1-1 to 2-10&lt;br&gt; <br /> <br /> pcr2 primer: 2-11 to 3-24&lt;br&gt; <br /> <br /> [[Image:1619PCR 5-17-2013.JPG]]&lt;br&gt; <br /> <br /> template: pcr1+pcr2&lt;br&gt; <br /> <br /> primer: 1-1 to 3-24&lt;br&gt; <br /> <br /> [[Image:1619+1739+1742+1748 clone 5-7-2013.JPG]] <br /> <br /> 1: vector pET28b &lt;br&gt; <br /> <br /> Nhe1 + Not1&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2013-5-23 <br /> <br /> Make miniprep and digest Nhe1 + Not1 &lt;br&gt; <br /> <br /> [[Image:1619 digest 5-23-2013.jpg]]&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 /> *&amp;lt;br&amp;gt;<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> == &#039;&#039;&#039;fix of Cloning of 1619-pET28b-T4DNAPol&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; fix of Cloning of 1519-pT7A3-T3-DNA-Ligase <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-6-18---2013-6-24 <br /> <br /> &#039;&#039;&#039;Notebook:0&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:1619&#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 /> template:1619-1 (above2013/5/1)&lt;br&gt; <br /> <br /> pcr1 primer: 1-1 to 2-10 <br /> <br /> pcr2 primer: 2-11 to 3-24&lt;br&gt; <br /> <br /> [[Image:1619 pcr1+pcr2 5-6-2013.JPG]]&lt;br&gt; <br /> <br /> template: pcr1+pcr2 <br /> <br /> primer: 1-1 to 3-24&lt;br&gt; <br /> <br /> [[Image:1619PCR 5-31-2013.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:1619clone 6-20-2013.JPG]]&lt;br&gt; <br /> <br /> &amp;nbsp;lane 1:digest PCR product &lt;br&gt; <br /> <br /> lane 2: pET28b &lt;br&gt; <br /> <br /> Nhe+Not1&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2013-6-24 <br /> <br /> Make miniprep and digest with&amp;nbsp; Nhe+Not1&lt;br&gt; <br /> <br /> [[Image:555+556+1619 digest 6-24-2013.JPG]] <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 /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> == &#039;&#039;&#039;fix of Cloning of 1619-pET28b-T4DNAPol&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; fix of Cloning of 1519-pT7A3-T3-DNA-Ligase <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-6-24---2013-6-27 <br /> <br /> &#039;&#039;&#039;Notebook:0&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:1619&#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 /> template:1619-1 (above2013/5/1)&lt;br&gt; <br /> <br /> pcr1 primer: 1-1 to 2-10 <br /> <br /> pcr2 primer: 2-11 to 3-24&lt;br&gt; <br /> <br /> [[Image:1619pcr1+2 6-24-2013.JPG]]&lt;br&gt; <br /> <br /> template: pcr1+pcr2 <br /> <br /> primer: 1-1 to 3-24&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:1619PCR 6-25-2013.JPG]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:1619clone 6-24-2013.JPG]]&lt;br&gt; <br /> <br /> &amp;nbsp;lane 3:digest PCR product &lt;br&gt; <br /> <br /> lane 4: pET28b <br /> <br /> Nhe+Not1&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2013-6-27 <br /> <br /> Make miniprep and digest with&amp;nbsp; Nhe+Not1&lt;br&gt; <br /> <br /> [[Image:1619digest 6-27-2013.JPG]]&lt;br&gt; <br /> <br /> Comment by Paul: do not use expensive enzyme such as Not1 for screening <br /> <br /> &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;fix of Cloning of 1619-pET28b-T4DNAPol&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; fix of Cloning of 1619-pT7A3-T3-DNA-Ligase <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2013-7<br /> <br /> &#039;&#039;&#039;Notebook:05846&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:1619&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039;Shanyi&lt;br&gt;<br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === cloning ===<br /> <br /> template:1619-1 (above2013/5/1)&lt;br&gt; <br /> <br /> pcr1 primer: 1-1 to 2-10 <br /> <br /> pcr2 primer: 2-11 to 3-24&lt;br&gt; <br /> <br /> [[Image:1619fixpcr.JPG‎ ]]&lt;br&gt; <br /> <br /> template: pcr1+pcr2 <br /> <br /> primer: 1-1 to 3-24&lt;br&gt; <br /> <br /> [[Image:1619fixpcr2.JPG‎ ]]<br /> <br /> digest with Nhe+Not1&lt;br&gt; <br /> <br /> [[Image:Digest_for_1779_1619.JPG‎ ]]<br /> <br /> &lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2013-6-27 <br /> <br /> Make miniprep and digest with HindIII&lt;br&gt; <br /> <br /> [[Image:1619diget.JPG‎]]<br /> <br /> &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 /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> [[Category:Cloning]]</div> Paul.xu