Editing 2436-pET30-30GFP

== '''Cloning of <span dir="auto">2436-pET30-30GFP</span>''' ==

'''Date:&nbsp;''' 2015-April

'''''Clone ID''': 2436''

'''Experiment''':&nbsp; Xin

'''Results''':

=== 1.&nbsp;<span class="mw-headline" id="Gene_Synthesis_by_PCR_using_oligo_as_template">Gene Synthesis </span>by two sections: ===

=== Section I: 1-1 __ 1-10; Section II: 1-11 __1-20 ===

[[File:2423, 2436 PCR 4-9-11.JPG|319x129px|RTENOTITLE]]





combine two sections together, PCR again.

[[File:2423 2436 PCR 4-9-15.JPG|276x122px|RTENOTITLE]]

&nbsp; 2. Clone: 

Insert the PCR product into vector 440. 

[[File:440 digestion.JPG|RTENOTITLE]]

Ligation and transformation.&nbsp; 

== '''''Screening''''':  ==
<div><span dir="auto">&nbsp;[[File:IMG 0317 2436 SCREENING1.JPG|285x162px|RTENOTITLE]]</span>&nbsp; The insectionthe size is about 1200bp, but should be about 850 bp.<br/></div><div><br/></div><div>Redigested again using different enzyme: XhoI/ NdeI<br/></div><div>[[File:2436-screening- II .JPG|271x186px|RTENOTITLE]]&nbsp; The size is still not right.<br/></div><div><br/></div><div>Selected one clony to sequence, the clone looks all right. The marker on the argrose gel isn't very accurcy.<br/></div><div><br/></div>
== '''Fix on April, 28, 2015''' ==

&nbsp;First PCR fragment: use the oligos 2436-30-GFP-1;2436-30-GFP-12 as primers

&nbsp;Second PCR fragment: use the oligos 2436-30-GFP-13;2436-30-GFP-20 as primers
<div>[[File:2423,2436,2421 PCR.JPG|337x141px|RTENOTITLE]]<br/></div><div><br/></div><div>Join two PCR together:<br/></div><div>[[File:2436, 2423, 2421 PCR.JPG|397x148px|RTENOTITLE]]<br/></div><div><br/></div><div>Screening: <br/></div><div>[[File:2434 2436 screening 5-1-15.JPG|462x148px|RTENOTITLE]]<br/></div><div><br/></div><div>All 8 colonies have right insert. <br/></div>
== '''Result intepretation/Summary''' ==

== '''Conclusion''' ==

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

== '''Conclusion''' ==
[[Category:Pages with broken file links]] [[Category:Cloning]]