Team:Groningen/Notebook/tasA Digestion t35
Blue Bio Energy
tasA Digestion (t35)
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 not much visible on gel. The experiment was repeated with more DNA.
Restriction
00:00, 11 June 2015 - 00:00, 11 June 2015
All tasA samples were digested to check for the insert. For this a Mastermix for digestion was made.
Component
Amount
10x buffer (2.1)
22 µL
EcoRI
9 µL
PstI
9 µL
\( \mathrm{H_2O}\)
167 µL
Mastermix for digestion.
Component
Amount
Mastermix
18 µL
DNA
2 µL
Sample for digestion.
The samples were incubated at 37 °C for 2 hours and 15 minutes. Afterwards they were put on a 1% agarose gel with 1:50000 DNA stain G. The gel was run on 120 V.
Sample:
5 µL sample
1 µL 6x buffer
Ladder:
10 µL Quick-Load® Purple 2-Log DNA Ladder (0.1 - 10.0 kb)
On the gel there was not much visible, so the procedure was repeated with more DNA.