https://eln.excellgen.com/index.php?action=history&feed=atom&title=998_pT7A3-6His-NLS-dCas9-NLS-6xHis 998 pT7A3-6His-NLS-dCas9-NLS-6xHis - Revision history 2026-10-18T12:55:49Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=998_pT7A3-6His-NLS-dCas9-NLS-6xHis&diff=18996&oldid=prev Crm at 17:39, 13 August 2018 2018-08-13T17:39:16Z <p></p> <p><b>New page</b></p><div>&#039;&#039;&#039;Vector digestion&#039;&#039;&#039;<br /> <br /> [[File:IMG 0858.JPG|RTENOTITLE]]<br /> <br /> 998 Vector was not digested completely<br /> <br /> &#039;&#039;&#039;PCR of 998_1000&#039;&#039;&#039;<br /> <br /> [[File:IMG 0851.JPG|RTENOTITLE]]<br /> <br /> &#039;&#039;&#039;Miniprep digestion of 998_1000_994_995&#039;&#039;&#039;<br /> <br /> {| border=&quot;1&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; style=&quot;width:500px;&quot;<br /> |-<br /> | #<br /> | Lane<br /> | Enzymes<br /> | Size<br /> | Note<br /> |-<br /> | 998<br /> | 1-4<br /> | HindIII+Xho1<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 1000<br /> | 5-8<br /> | HIndIII+Xho1<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 994<br /> | 9-12<br /> | BamH1+Xho1<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> |-<br /> | 995<br /> | 13-16<br /> | BamH1+Xho1<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> |}<br /> <br /> &#039;&#039;&#039;&#039;&#039;[[File:IMG 0873.JPG|RTENOTITLE]]&#039;&#039;&#039;&#039;&#039;<br /> <br /> <br /> <br /> <br /> == &#039;&#039;&#039;Cloning of 998 pT7A3-6His-NLS-dCas9-NLS-6xHis&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 998 pT7A3-6His-NLS-dCas9-NLS-6xHis<br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;06, 27, 2018<br /> <br /> &#039;&#039;&#039;Notebook: 008840&#039;&#039;&#039;<br /> <br /> &#039;&#039;&#039;Page:008958&#039;&#039;&#039;<br /> <br /> &#039;&#039;&#039;Clone ID: 998&#039;&#039;&#039;<br /> <br /> &#039;&#039;&#039;Experimenters: &#039;&#039;&#039;yijia<br /> <br /> === Cloning ===<br /> <br /> &#039;&#039;&#039;998 PCR&#039;&#039;&#039;<br /> <br /> [[File:IMG 1027.JPG|RTENOTITLE]]<br /> <br /> <br /> <br /> &#039;&#039;&#039;Miniprep of 3206_995_998&#039;&#039;&#039;<br /> <br /> [[File:IMG 1092.JPG|RTENOTITLE]]<br /> <br /> &#039;&#039;&#039;Miniprep digestion of 3218_998&#039;&#039;&#039;<br /> <br /> {| border=&quot;1&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; style=&quot;width:500px;&quot;<br /> |-<br /> | #<br /> | lane<br /> | Enzymes<br /> | Size<br /> | Result<br /> |-<br /> | 998<br /> | 9-12<br /> | HindIII<br /> | &lt;br/&gt;<br /> | <br /> |-<br /> | &lt;br/&gt;<br /> | 13-16<br /> | HindIII+Xho1<br /> | &lt;br/&gt;<br /> | &lt;br/&gt;<br /> |}<br /> <br /> [[File:IMG 1101.JPG|RTENOTITLE]]<br /> <br /> == redo ==<br /> <br /> &#039;&#039;&#039;998 PCR&#039;&#039;&#039;<br /> <br /> {| border=&quot;1&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; style=&quot;width:500px;&quot;<br /> |-<br /> | #<br /> | Lane<br /> | style=&quot;width: 181px;&quot; | Enzymes<br /> | style=&quot;width: 209px;&quot; | Buffer<br /> |-<br /> | Kappa<br /> | 1,2<br /> | style=&quot;width: 181px;&quot; | <br /> Kappa HiFi<br /> <br /> | style=&quot;width: 209px;&quot; | <br /> 5xKappa buffer<br /> <br /> |-<br /> | Takara<br /> | 3,4<br /> | style=&quot;width: 181px;&quot; | Takara DNA polymerase<br /> | style=&quot;width: 209px;&quot; | 5xTakara buffer<br /> |-<br /> | Q5<br /> | 5,6<br /> | style=&quot;width: 181px;&quot; | Q5 HiFi<br /> | style=&quot;width: 209px;&quot; | 5xphusiob HF hot start buffer<br /> |}<br /> <br /> [[File:IMG 1124.JPG|RTENOTITLE]]<br /> <br /> &#039;&#039;&#039;redo PCR&#039;&#039;&#039;<br /> <br /> [[File:IMG 1182.JPG|RTENOTITLE]]<br /> <br /> == Date:8/2/2018 ==<br /> <br /> &#039;&#039;&#039;vector digestion of 3206_998&#039;&#039;&#039;<br /> <br /> [[File:IMG 1259.JPG|RTENOTITLE]]<br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === &amp;nbsp;Colony screen ===<br /> == Date:8/7/2018 ==<br /> <br /> &#039;&#039;&#039;Miniprep digestion of 998_931-1_931-1pjet1.2_129&#039;&#039;&#039;<br /> <br /> [[File:IMG 12720.JPG|RTENOTITLE]]<br /> <br /> &#039;&#039;&#039;Miniprep digestion 998&#039;&#039;&#039;<br /> <br /> &#039;&#039;&#039;1-3:HindIII+Xho1&#039;&#039;&#039;<br /> <br /> &#039;&#039;&#039;#&amp;nbsp; &amp;nbsp; &amp;nbsp; Ends&amp;nbsp; &amp;nbsp; &amp;nbsp; &amp;nbsp; &amp;nbsp; Length (bp)&#039;&#039;&#039;<br /> <br /> 1 HindIII-HindIII&amp;nbsp; &amp;nbsp; &amp;nbsp;2345<br /> <br /> 2 HindIII-HindIII&amp;nbsp; &amp;nbsp; &amp;nbsp;1034<br /> <br /> 3 HindIII-XhoI&amp;nbsp; &amp;nbsp; &amp;nbsp; &amp;nbsp; &amp;nbsp;774<br /> <br /> 4 XhoI-HindIII &amp;nbsp; &amp;nbsp; &amp;nbsp; &amp;nbsp; 118<br /> <br /> &#039;&#039;&#039;4-6:Nhe1+Kpn1 679bp&#039;&#039;&#039;<br /> <br /> [[File:IMG 1273.JPG|RTENOTITLE]]<br /> == Date:8/8/2018 ==<br /> <br /> &#039;&#039;&#039;miniprep digstion 998&#039;&#039;&#039;<br /> <br /> [[File:IMG 1279.JPG|RTENOTITLE]]<br /> <br /> &#039;&#039;&#039;choose #1,#3 to do another digestion&#039;&#039;&#039;<br /> <br /> &#039;&#039;&#039;Miniprep digestion of #1,#3&#039;&#039;&#039;<br /> <br /> [[File:IMG 1280.JPG|RTENOTITLE]]<br /> <br /> ==Date:8/13/2018==<br /> redo<br /> &#039;&#039;&#039;vector digstion&#039;&#039;&#039;<br /> <br /> [[File:IMG 1285.JPG]]<br /> <br /> &#039;&#039;&#039;998 PCR Screening&#039;&#039;&#039;<br /> <br /> [[File:IMG 1289.JPG]]<br /> <br /> &#039;&#039;&#039;998 miniprep 1&#039;&#039;&#039;<br /> <br /> [[File:IMG 1296.JPG]]<br /> <br /> &#039;&#039;&#039;998 Miniprep 2&#039;&#039;&#039;<br /> <br /> [[File:IMG 1298.JPG]]<br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> [[Category:Cloning]]</div> Crm