June 19
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* Transformants were checked and no growth found in -ve but in j22261,j22271,j22281,j22301. | * 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:''' | '''Microscopy:''' | ||
| + | |||
| + | * pLac yfp imaging done after 12,14 and 16 hrs. | ||
| + | |||
| + | * pLac cfp imaging done after 12 hrs. | ||
| + | |||
| + | |||
| + | '''Analysis''' | ||
Revision as of 09:08, 28 June 2007
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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
