Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/09/17

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12PM Pulled RhlI out at 12pm. pH looks between 7 and 8 (paper strip)


Spun 1mL of cells, resuspended in 100ul water, checked GFP: 158.
Split cells and LB into four-2mL tubes
Spun at 12.0g for 3min
Transferred 1mL LB supernatant to new tubes

One-2mL tube for later induction (stored at -20)
Three-15mL conical tubes

Added 1mL ethyl acetate to LB

Vortex for 15 seconds, sit for 2min
Repeat 5 times
Fifth time,let sit for 10 minutes

Transfer 1mL of the top layer to 1.5mL tube Let sit for 15-30 seconds Transfer top layer to new 15mL conical tube Store at -20°C

3PM Pulled Rhl2 out

OD650GFP in LBGFP in H20pH

Repeated extraction procedure

6PM Pulled Rhl3 out

OD650GFP in LBGFP in H20pH

Repeated the extraction procedure for Rhl3

Tranformations into DH5αT and BL21

  1. J27 (F2620-RFP)
  2. RhlI 8/31/13
  3. EsaI sender A3
  4. GFP-Lux rec 1

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