Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/09/25

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mm/dd/yyyy

Sender/ReceiverOD650GFPRFPpH
EsaI 1 0.6971034 7-8
EsaI 0.6781023 7-8
RhlI 1 0.7076561 7-8
RhlI 0.7197153 7-8
Ctrl 1 0.7375494 7-8
Ctrl 0.6925273 7-8
LuxR-GFP 0.5491510
LuxR-RFP 0.536 748


Spun all cultures in 2mL tubes for 3min at 12.9RPM (15.4g). Moved sender sup to 15mL conical tubes (combined all 10mLs). 3mL each sender into culture tubes.

Resuspended receivers into 150ul plain LB. moved 50ul each into 3mL each sender Started shaking at 5PM

Extracted remaining 3mLs into 1mL ethyl acetate

15s vortex, 2min wait 5 times. 10 min wait last time

still didn't separate (all balled up on top of the media). added a second 1mL of ethyl acetate, repeated extraction protocol to get 1mL


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