Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/03/21
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Picked 2 E0240 colonies into 2ml. Checked the plates that I pipetted S-R pairs onto last night under UV, didn't see anything. Pipetted the co-cultures onto the plate (15-20ul). Didn't see anything. Miniprepped at 6ish. low yields. Cut E0240 with X/P, ran on gel.
Grew from 11am-6 or 7pm. Spun down co-cultures at 12.5ish for 2mins. Pours off supernatant, resuspended in ~100ul H2) (ended up being ~150 total with residual LB and cells). Used plate reader to check fluorescence for CFP and got higher readings for RhlI-RhlR.
Transformed into 50ul of BL21. At RT for 10min, ice for 13min, 42deg for 45s
4-6ml cultures of senders 3-35ml cultures of RhlR Started at 11:30PM