Team:Groningen/Notebook/tasA Digestion t36
Blue Bio Energy
tasA Digestion (t36)
tasA plasmid was digested to check for 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.
There was nothing visible on gel. The gel was probably too hot when adding the DNA stain G.
Restriction
00:00, 15 June 2015 - 00:00, 15 June 2015
Digested tasA from backbone using EcoRI and PstI to check ligation. For this, a Mastermix was made for 13 samples (∼300 µL).
Component
Amount
10x buffer (2.1)
30 µL
EcoRI
10 µL
PstI
10 µL
\( \mathrm{H_2O}\)
185 µL
Mastermix digestion.
Component
Amount
DNA
5 µL
Mastermix
15 µL
Components per digestion sample.
The samples were incubated for 2 hours at 37 °C. Afterwards they were loaded on 1% agarose gel with 1:50000 DNA stain G.
Sample:
20 µL digestion sample
4 µL 6x buffer
Ladder:
10 µL Quick-Load® Purple 2-Log DNA Ladder (0.1 - 10.0 kb)
There was nothing visible on the gel. The gel was probably too hot when adding the DNA stain G.