Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2014/05/21

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Golden gate 1:10 RhlR and EsaR receivers grew but there were colonies on the backbone plate

Ligation Colonies
Bb13
RhlR23
EsaR14



Ran PCR, extension time 3:30 at 50°C

Template F primer R primer
pTet-mCh13097
LuxI GFP rec131132
LuxR 1:10 colony 1103104
LuxR 1:10 colony 2103104
LuxR 1:10 colony 3103104
LuxR 1:10 colony 4103104
LuxR 1:10 colony 5103104
LuxR 1:10 colony 5103107


Ran above PCR on gel

Template F primer R primer Expected band size Actual band size
pTet-mCh1309731303130
LuxI GFP rec131132610no band
LuxR 1:10 colony 11031141817no band
LuxR 1:10 colony 210311418171817
LuxR 1:10 colony 31031141817no band
LuxR 1:10 colony 41031141817no band
LuxR 1:10 colony 510311418171817
LuxR 1:10 colony 510310710351035



picked LuxI and RhlI GFP senders into 4mLs media and shook overnight to screen for successful receiver transformations.



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