Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/09/19

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==09/19/2013==
RhlI
RhlI
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09/19/2013

RhlI

OD650GFP in LBpH
0.61299297-8

EsaI

OD650GFP in LBpH
0.5512577-8

Double checked super weak EsaI GFP expression and it's correct. Pelleted cells were not as green as RhlI.
F2620-RFP

OD650mCherry in LB
0.544668

F2620-GFP

OD650GFP in LB
0.5021166


Control supernatant was DH5αT K092900-E0420 which was not a perfect control. pH was 8-9.
11:40am
measured mCherry Ex:572/Em:607 Spun Rhl and Esa at 10:30am. Moved 1mL to round bottom tubes. Added 40uL LuxR-GFP or LuxR-RFP. Added 1ul amp. RhlI

SenderReceiver
RhlILuxR-GFP
EsaILuxR-GFP
CtrlLuxR-GFP
RhlILuxR-RFP
EsaILuxR-RFP
CtrlLuxR-RFP


Thawed 1mL supernatant tubes, transferred to round bottom tubes. Added 40uL LuxR-GFP. Added 1uL amp.

SenderReceiver
RhlI 1LuxR-GFP
RhlI 2LuxR-GFP
RhlI 3LuxR-GFP



minipreps picked 9/18/13

PartResult
K081002grew slowly
I13522-3grew slowly
K092900-E0420 (80.5)didn't grow
F2620-GFPgrew
RhlIgrew
F2620-RFPgrew
K081020grew
EsaIgrew
K092900-E0420 (80)grew


pretty sure i put these in the freezer and did them another day.


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