Team:Groningen/Notebook/tasA Colony PCR t102
Blue Bio Energy
tasA Colony PCR (t102)
A colony PCR was performed to check for the right insert.
Our goal is to overexpress TasA by adding a second copy of the tasA gene under the control of a salt inducible promoter. This overexpression will lead to a stronger biofilm when the biofilm is exposed to salt water.
Four colonies were PCR'ed and grown overnight.
Colony PCR
00:00, 18 August 2015 - 00:00, 18 August 2015
Plates were checked for colonies. Plate with salt promoter in pSB1C3 contained colonies. 4 colonies were checked with colony PCR, therefore VF and VF2 primers were used.
#
Temperature
Time
Step
1
98 °C
5:00
Initial denaturation
2
98 °C
0:10
Denaturation
75 °C
0:30
Annealing
72 °C
1:30
Extension
repeat 35x
3
72 °C
5:00
Final extension
PCR Thermocycle
Each transformation had at least one correct insert according to the PCR. This colony was grown in LB with corresponding antibiotics overnight at 37 °C.