Editing 1292 pET30b-PTD-DRBD-sso7d (section)

=== Cloning<br>  ===

Date:&nbsp;August 1, 2012 

[[Image:1292 Amp.JPG]] 

Synthesized template via gene synthesize, expected size ~400 bp 

Cut out band in lane 2, purify 

[[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; === 

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 

Date:&nbsp;August 2, 2012<br> 

Digested 368 vector (lane 3) with RE Kpn1 and Nde1, ligate together with digested insert from yesterday<br> 

[[Image:1292 Vector.JPG]] 

Ligate together over night, transform tomorrow morning 

Date:&nbsp;August 13, 2012<br> 

Screening came back mostly negative<br> 

Synthesized template via gene synthesize, expected size ~400 bp 

Cut out band in lane 2, purify <br> 

[[Image:1292 0813 Amp.JPG]] 

Digested insert (see lane 3) and 368 vector (lane 4) with RE Kpn1 and Nde1, cut out and purify. LIgate together overnight<br> 

[[Image:1292 0813 Digestion.JPG]]