A variety of ligations were carried out last week and to test there success we carried out single colony PCR using Psb primers to verify if an insert is present in the vector AK3. For each ligation thress seperate colonies were picked to carry out SCP and a negative colony where no DNA was present. The gel below shows the resuls of this experiment:
The conditions used were as follows:
1 cycle - 95oC for 30 seconds
30 cycles - 95oC for 30 seconds, 60oC for 30 seconds,72oC for 30 seconds
1 cycle - 72oC for 2 minutes
The numbers of the ligations correspond to the following ligation reactions:
5-Eps+AK3
8-PgsiB-gsiB+AK3
9-PgsiB-spoVG+AK3
12-Pveg-spoVG
Dry Lab
Tracked a total ot 90 cells. Run velocity profile is shown below: