Team:TU-Delft/Notebook/2013/09/14/

1. Did Colony PCR on :









2. The pT7 Lysis was run again with a Control to double check:



3. Transformation was carried out on:
          pBAD Ulp TT pT7 SUMO Magainin Col 2 --> BL21
          pBAD Ulp TT pT7 SUMO Maximin H5 Col 5 --> BL21

4. The lysis experiment was carried out to check whether on inducing the pT7 promoter with IPTG, the cells lyse. The over night grown culture was diluted 1/50 around 12:00 Noon. The OD was monitored till it reached 0.5 which was around 3.30 PM.
Then 30 mL of culture was taken in three different tubes and induced with 10mM, 1mM and 0.1mM concentrations.
The OD was monitored to check for death of cells :
          Time           --> 10mM           1mM           0.1mM
          At 4.30 PM           --> 0.77           0.78           0.83
          At 5.15 PM           --> 0.91           0.93           0.94
          At 6.30 PM           --> 1.05           1.05           1.14

The experiment proved that the lysis of cells was not happening. This may be due to a wrong plasmid while transformations or the BL21 strain was not correct.
So we started afresh to transform another pT7 Lysis from a different colony into BL21 plysS strain.