Difference between revisions of "Team:Paris Saclay/Notebook/July/8"

(→‎Colony screening PCR on bacteria transformed with pPS16_004 and pPS16_007)
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===Visualization===
 
===Visualization===
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====Colony screening PCR on bacteria transformed with [[Team:Paris_Saclay/Notebook/June/28#pPS16_004|pPS16_004]] and [[Team:Paris_Saclay/Notebook/June/28#pPS16_007|pPS16_007]]====
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====Colony screening PCR<div id="Colony screening PCR"></div> on bacteria transformed with [[Team:Paris_Saclay/Notebook/June/28#pPS16_004|pPS16_004]] and [[Team:Paris_Saclay/Notebook/June/28#pPS16_007|pPS16_007]]====
 
''By Alice''
 
''By Alice''
  

Revision as of 15:29, 12 July 2016

Tuesday 8th July

Lab work

Visualization

Colony screening PCR
on bacteria transformed with pPS16_004 and pPS16_007

By Alice

After transformation, only white bacteria are selected (blue white screen). They are expected to have plasmids with inserts of interest. PCR with DreamTaq DNA Polymerase is performed following this protocol. Clones selected for PCR are kept on LB+Amp+XGal(2µLXGal/1000µLH2O)+IPTG(2µLIPTG/1000µLH2O). Two primers (1152_pheoF and 1151 5_pheoR) upstream and downstream the insertion site are chosen. . PCR products expected below :

Plasmids Band size (bp)
pPS16_004 846
pPS16_007 744

No PCR products are observed, due to an error in the protocol : MilliQ water was used instead of steril water. The same PCR will be performed on 11/07/16.