Team:Groningen/Notebook/tasA Digestion t61
Blue Bio Energy
tasA Digestion (t61)
PliaG and tasA (T2) were digested to make the 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 of PliaG and T2 was carried out.
Restriction
13:22, 6 August 2015 - 18:40, 6 August 2015
PliaG was digested for ligation with T2. Restriction enzymes were used that create a scar. For the digestion the restriction enzymes EcoRI and SpeI were used for the promoter and XbaI and EcoRI for T2. Below are listed the amounts that were added of each component.
#
Component
Amount
1
PliaG DNA
5 µL
10x buffer
2 µL
EcoRI
1 µL
SpeI
1 µL
\( \mathrm{H_2O}\)
11 µL
2
T2 DNA
5 µL
10x buffer
2 µL
XbaI
1 µL
EcoRI
1 µL
\( \mathrm{H_2O}\)
11 µL
Components for digestion.
The sample was digested for 2 hours at 37 °C.