Editing 1292 pET30b-PTD-DRBD-sso7d

== '''Cloning of 1292 pET30b-PTD-DRBD-sso7d'''  ==

'''Purpose:''' Cloning of 1292 pET30b-PTD-DRBD-sso7d 

'''Date: '''August 1, 2012<br> 

'''Notebook:&nbsp;''' 

'''Page:''' 

'''Clone ID: '''1292 

'''Experimenters: '''Jennifer Truong 

== '''Results'''  ==

=== 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]] 

=== <u></u>Colony Screening<br>  ===

Date:&nbsp;August 7, 2012<br> 

[[Image:1292 0807 Screening.JPG]] 

Digested MiniPrep with RE&nbsp;Kpn1 and Nde1 

4 and 5 may be correct, maybe will digest again 

[[Image:1292 0807 Screening 2.JPG]] 

Digested sample 4 (lane 1)&nbsp;and sample 5 (lane 2)&nbsp;again with RE&nbsp;Nde1 and Kpn1 

Very faint band at expected size (~400)&nbsp; 

Send sample 5 to sequencing 

Date:&nbsp;August 20, 2012 

----

[[Image:1292 0820 Screening.JPG]] 

Cannot tell, bad digestion 

<br> Date:&nbsp;OCO 26, 2012 

Digested insert&nbsp; and&nbsp; vector&nbsp; with&nbsp; Nde1 + Xho1<br> 

[[Image:1290V15 1292368 1293V15 1352 13041280DIGEST 10 26 2012.JPG]]<br> 

Ligate , transform&nbsp; 

<br> 

=== <u></u>Colony Screening<br>  ===

[[Image:1292 1307DIGEST10 31 2012.JPG]]<br> 

sent to sequence miss one g<br>

PCR<br>

[[Image:1136_1139_1292PCR.JPG]]<br>

= 1117PCR-1118PCR 1422 1277 V15 1292 368 1139 1141 1147.JPG.JPG =

<br>

DIGEST pET30b-sso7d-KX 368 WITH Nde1 + Kpn1<br>

[[Image:1117PCR-1118PCR 1422 1277 V15 1292 368 1139 1141 1147.JPG.JPG]]

SCREENING
 [[Image:1481 screening 2012 12 7.jpg]]<br> 


== '''Result intepretation/Summary'''  ==

<br> 

<br> 

== '''Conclusion'''  ==

<br> 

[[Category:Cloning]]