From 2007.igem.org

Back to Bangalore

Back to e-Notebook

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