Team:Groningen/Notebook/tasA PCR purification t70
Blue Bio Energy
tasA Gel purification (t70)
The digestion product was purified to get rid of the restriction enzymes.
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 tasA and PliaG DNA was gel purified.
Gel purification
10:30, 13 August 2015 - 20:15, 13 August 2015
Afterwards they were loaded on a 1% agarose gel with DNA stain G 1:50000. For the DNA samples 20 µL was used with 4 µL 6x buffer. As a ladder 1 µL of the GeneRuler™ 1 kb DNA Ladder was used. The gel was run for ∼45 minutes at 100 V. The bands with the right sizes were cut from the gel and dissolved in twice as much elution buffer. The GeneJet PCR purification kit was used to extract the DNA, using 21µL elution buffer. With Nanodrop the concentrations were determined.
Sample
Concentration
1
4.0 ng/µL
2
5.1 ng/µL
Concentrations of samples determined with Nanodrop.