Team:Groningen/Notebook/tasA Miniprep t67
Blue Bio Energy
tasA Miniprep (t67)
Extract plasmid from E. coli.
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 miniprep of the E. coli transformed with tasA and PliaG was performed.
Plasmid DNA purification
12:00, 11 August 2015 - 15:30, 11 August 2015
The cultures of sample 3 and 7 were miniprepped and loaded on 1% agarose gel with DNA stain G 1:50000. For each DNA sample 1 µL was added to the gel, with 4 µL \( \mathrm{H_2O}\) and 1 µL 6x buffer. As a ladder 10 µL of the Quick-Load® Purple 2-Log DNA Ladder (0.1 - 10.0 kb) was used. The gel was run for ∼30 minutes at 100 V. On the gel afterwards nothing was visible, so the DNA concentration needed to be checked again.
10:30, 13 August 2015 - 20:15, 13 August 2015
The concentrations of the samples 3 and 7 were checked again and were found to be the following.
Sample
Concentration
3
3.3 ng/µL
7
9.3 ng/µL
Concentrations of samples determined with Nanodrop.