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1292 pET30b-PTD-DRBD-sso7d - Revision history
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Paul.xu at 15:33, 12 December 2012
2012-12-12T15:33:06Z
<p></p>
<p><b>New page</b></p><div>== '''Cloning of 1292 pET30b-PTD-DRBD-sso7d''' ==<br />
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'''Purpose:''' Cloning of 1292 pET30b-PTD-DRBD-sso7d <br />
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'''Date: '''August 1, 2012<br> <br />
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'''Notebook:&nbsp;''' <br />
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'''Page:''' <br />
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'''Clone ID: '''1292 <br />
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'''Experimenters: '''Jennifer Truong <br />
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== '''Results''' ==<br />
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=== Cloning<br> ===<br />
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Date:&nbsp;August 1, 2012 <br />
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[[Image:1292 Amp.JPG]] <br />
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Synthesized template via gene synthesize, expected size ~400 bp <br />
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Cut out band in lane 2, purify <br />
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[[Image:1292 Digestion Insert.JPG]]Ligate , transform&amp;nbsp; <br><br>&lt;br&gt; <br><br>&lt;br&gt; <br><br>=== &lt;u&gt;&lt;/u&gt;Colony Screening&lt;br&gt;&nbsp; === <br />
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Digested insert (see lane 3)&nbsp;and 368 vector (lane 4)&nbsp;with RE Kpn1 and Nde1, vector has too band bands, digest again, save insert for ligation over night <br />
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Date:&nbsp;August 2, 2012<br> <br />
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Digested 368 vector (lane 3) with RE Kpn1 and Nde1, ligate together with digested insert from yesterday<br> <br />
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[[Image:1292 Vector.JPG]] <br />
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Ligate together over night, transform tomorrow morning <br />
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Date:&nbsp;August 13, 2012<br> <br />
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Screening came back mostly negative<br> <br />
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Synthesized template via gene synthesize, expected size ~400 bp <br />
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Cut out band in lane 2, purify <br> <br />
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[[Image:1292 0813 Amp.JPG]] <br />
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Digested insert (see lane 3) and 368 vector (lane 4) with RE Kpn1 and Nde1, cut out and purify. LIgate together overnight<br> <br />
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[[Image:1292 0813 Digestion.JPG]] <br />
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=== <u></u>Colony Screening<br> ===<br />
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Date:&nbsp;August 7, 2012<br> <br />
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[[Image:1292 0807 Screening.JPG]] <br />
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Digested MiniPrep with RE&nbsp;Kpn1 and Nde1 <br />
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4 and 5 may be correct, maybe will digest again <br />
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[[Image:1292 0807 Screening 2.JPG]] <br />
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Digested sample 4 (lane 1)&nbsp;and sample 5 (lane 2)&nbsp;again with RE&nbsp;Nde1 and Kpn1 <br />
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Very faint band at expected size (~400)&nbsp; <br />
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Send sample 5 to sequencing <br />
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Date:&nbsp;August 20, 2012 <br />
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[[Image:1292 0820 Screening.JPG]] <br />
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Cannot tell, bad digestion <br />
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<br> Date:&nbsp;OCO 26, 2012 <br />
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Digested insert&nbsp; and&nbsp; vector&nbsp; with&nbsp; Nde1 + Xho1<br> <br />
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[[Image:1290V15 1292368 1293V15 1352 13041280DIGEST 10 26 2012.JPG]]<br> <br />
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Ligate , transform&nbsp; <br />
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=== <u></u>Colony Screening<br> ===<br />
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[[Image:1292 1307DIGEST10 31 2012.JPG]]<br> <br />
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sent to sequence miss one g<br><br />
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PCR<br><br />
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[[Image:1136_1139_1292PCR.JPG]]<br><br />
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= 1117PCR-1118PCR 1422 1277 V15 1292 368 1139 1141 1147.JPG.JPG =<br />
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DIGEST pET30b-sso7d-KX 368 WITH Nde1 + Kpn1<br><br />
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[[Image:1117PCR-1118PCR 1422 1277 V15 1292 368 1139 1141 1147.JPG.JPG]]<br />
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SCREENING<br />
[[Image:1481 screening 2012 12 7.jpg]]<br> <br />
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== '''Result intepretation/Summary''' ==<br />
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== '''Conclusion''' ==<br />
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[[Category:Cloning]]</div>
Paul.xu