Team:Groningen/Notebook/tasA PCR purification t63
Blue Bio Energy
tasA Gel purification (t63)
T2 (tasA) and PliaG were gel purified to be used in a ligation afterwards.
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 T2 (tasA) DNA and the PliaG promoter were gel purified.
Gel purification
13:22, 6 August 2015 - 18:40, 6 August 2015
The digested samples (T2 and PliaG) were loaded on 1% agarose gel with DNA stain G 1:50000. On the gel were loaded 20 µL of the sample DNA with 4 µL buffer. As a ladder 10 µL of the 2log NEB ladder was used. The gel was run for ∼30 minutes at 100 V. The right bands were cut from the gel and twice as much binding buffer was added. The samples were purificated using the Thermo Scientific GeneJET PCR Purification Kit and their concentrations were checked using Nanodrop.
Sample
Concentration
PliaG dig
4.6 ng/µL
T2 dig
4.9 ng/µL
Concentrations of samples determined with Nanodrop.