Team:Groningen/Notebook/tasA Digestion t56
Blue Bio Energy
tasA, Pveg and Psalt Digestion (t56)
tasA, the salt promoter and the Pveg promoter were digested to make ligation possible.
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.
The digestion succeeded, the right bands were cut out of gel.
Restriction
09:30, 28 July 2015 - 16:00, 28 July 2015
The digestion of the T2 sample of 10 July with two promoters was performed, both with the Pveg promoter from the iGEM distribution kit 2015 (plate 1, 20G) and the Psalt promoter. For the promoters EcoRI and SpeI were used as restriction enzymes, and XbaI and EcoRI for the backbone. The following samples were made.
#
Component
Amount
1
T2 DNA
5 µL
10x buffer
2 µL
EcoRI
1 µL
XbaI
1 µL
\( \mathrm{H_2O}\)
11 µL
2
Pveg DNA
10 µL
10x buffer
2 µL
EcoRI
1 µL
SpeI
1 µL
\( \mathrm{H_2O}\)
6 µL
4
Psalt DNA
5 µL
10x buffer
2 µL
EcoRI
1 µL
SpeI
1 µL
\( \mathrm{H_2O}\)
11 µL
Components digestion samples.
The samples were digested at 37 °C for 2 hours.