June 19

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Experiments

  • Transformants were checked and no growth found in -ve but in j22261,j22271,j22281,j22301.
  • Single colony streaking of transformed colonies was done.
  • pT luxI cfp inoculated in 3ml LB+3ul amp with K12Z1 in 3ml LB+3ul spec for control.
  • pLac yfp inoculated in 3ml LB+3ul amp with K12Z1 in 3ml LB+3ul spec for control.
  • OD measurement done with 12 small tubes with conc 0.01,0.1,1,3 ul cells after 10 hours in LB (triplets).
  • pT luxI cfp + K12Z1 induced with aTc (0,1,10,25,50,100 ng/ml in triplets for microscopy.
  • pT luxI cfp + K12Z1 induced with aTc (0,1,10,25,50,100 ng/ml in doublets for FACS.
  • pLac yfp + K12Z1 induced with IPTG (0,1,10,50,100,1000 uM).
  • FACS Recalibration carried out in afternoon.


Microscopy:

  • pLac yfp imaging done after 12,14 and 16 hrs.
  • pLac cfp imaging done after 12 hrs.


Analysis