Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/05/30
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BL21 sender cultures grew in 6mls and 2mls. RhlR-GFP still didn't grow.
Spun down 4ml of culture in 2-2ml tubes for each sender. Poured supernatant into round bottom tubes. Dropped some supernatant onto pH strip. Added 1ml of receiver cells to a 2ml centrifuge tube. Added 20ul of Amp and shook. Added 100ul of Receivers + (~4ul) Amp to each 4ml volume of Sender cells in round bottom tubes. Added 1ml of LB+Amp to add nutrients and lower the pH a little. Same Sender-Receiver pairs as above. Started shaking ~10:30AM.
Spun 1ml of supernatant transfer experiments and co-culture experiments in 2ml tubes at full speed for 3 minutes. Poured off supernatant, resuspended cells in 100ul water. Could see that some of the cell pellets were green while controls were not. Moved resuspended cells to plate for fluorescence reading. Added 100ul of each supernatant culture to plate as well for cell density readings.