Team:Groningen/Notebook/tasA Digestion t69
Blue Bio Energy
tasA Digestion (t69)
The digestion of tasA DNA and the PliaG promoter was performed to make the next step, ligation of the two part, 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.
tasA and PliaG were digested for ligation.
Restriction
10:30, 13 August 2015 - 20:15, 13 August 2015
The digestion was carried out using for the promoter PliaG, the restriction enzymes SpeI and PstI, and for tasA (T2), the enzymes XbaI and PstI. The following digestion samples were prepared.
#
Component
Amount
1
PliaG DNA
10 µL
10x buffer (2.1)
2 µL
SpeI
1 µL
PstI
1 µL
\( \mathrm{H_2O}\)
6 µL
2
T2 DNA
10 µL
10x buffer (2.1)
2 µL
XbaI
1 µL
PstI
1 µL
\( \mathrm{H_2O}\)
6 µL
Components for digestion.
The samples were incubated at 37 °C for 3.5 hours.