Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/03/23

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R-S didn't grow but R-only did. Picked R-colonies into S-R cultures. Added 1ml fresh LB in case the problem was old media, might need to check pH.
4PM: still hadn't grown much, but definitely growing.
9PM: still not growing well. decided to spin all 6mls and just take one time point.

1-34-67-910-12
LuxI 1-3LasI 1-3 RhlI 1-3 RhlR-only 1-3

Used 100ul to 1ml of LB to test OD. Added 1ml H2), flicked tube, spun, poured off H2), added 100ul more.

R1-3~150ul
C1~300ul (only 200ul went into the plate
C2~150Ul
C3~200ul


Looks like fluorescence corresponds to cell density. E. coli auto fluoresce in the same range than CFP is excited. Tried a range of wavelengths but was not able to get lower emissions:

Excitation (nM) Emission (nm)
439476
475500
450500
400500
400475
375476



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