Team:Groningen/Notebook/tasA PCR purification t57
Blue Bio Energy
tasA Gel purification (t57)
The right pieces of DNA were obtained by cutting them from the gel and purifying them.
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 digested samples were gel purified.
Gel purification
09:30, 28 July 2015 - 16:00, 28 July 2015
Afterwards a 1% agarose gel with stain G 1:50000 was run on 100 V for ∼45 minutes. 20 µL of DNA samples were mixed with 4 µL 6x buffer. As a ladder the 10 µL 2log NEB ladder was used. The right bands were cut out, weighed and dissolved in twice as much binding buffer. The samples were purified with the PCR purification kit (Genejet #K0701, lot 00265976) using 25 µL elution buffer. The concentrations were determined with Nanodrop.
Sample
Concentration
T2 dig (2)
4.5 ng/µL
Pveg (3)
1.4 ng/µL
Psalt (4)
5.0 ng/µL
Concentrations of samples determined by Nanodrop.