July 3
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Experiments
- pT.LuxI.C imaging was done as part of equivalence expt.
- pL LuxR.Y imaging was done as part of the eqiuvalence expt.
- Open loop experiment [E]
i) Cells were transferred to 50 mL Glu M9 in 250 mL flask with [aTc]=50 ng/mL and inoculum=1,0.1 uL/mL.
ii) pL.LuxR.Y.pR.C and K12Z1 were inoculated in LB.
iii) K12z1 was inoculated in Glu M9 and atc (1000)
iv) The contents of the flask with required OD (50 ng/mL) were filtered.
v) Seven dilutions for pL.LuxR.Y.pR.C were made with 50 old:50 new 2xM9 along with -ve control: K12z1
- The following -ve controls were also done :
i) pL.LuxR.Y.pR.C with K12Z1 grown medium,500 uM IPTG.
ii) pL.LuxR.Y.pR.C with new M9,500 uM IPTG.
- Open loop experiment [F]
i) Cells were transferred to 50 mL Glu M9 in 250 mL flask with [aTc]=0 ng/mL and inoculum=1,0.1 uL/mL
ii) pL.LuxR.Y.pR.C, K12Z1 and pL.YFP were inoculated in LB
iii) The contents of the flask with required OD ( ) were filtered.
