Team:Groningen/Notebook/Transformation a27
Blue Bio Energy
Transformation_a27
Transform Bacillus subtilis 3610 delta comI strain with purified full genomic DNA of BD 1807 strain.
To knock out abrB in Bacillus subtilis strains to obtain a more rigid biofilm. AbrB suppresses biofilm formation. Knocking out AbrB should result in a more rigid biofilm.
Transformation failed, as no colonies were found on the plate.
Transformation
07:07, 26 July 2015 - 00:00, 26 July 2015
B.s. 3610 comI colony was placed in MC according to protocol for Bacillus subtilis transformation. The tube was put in the shaking incubator at 37 °C.
At 12:22 the OD at 600 nm was measured of the transformation culture:0.00. The sample was put back in the shaking incubator at 37 °C for overnight growth.
11:15, 27 July 2015 - 00:00, 27 July 2015
The OD at 600 nm was measured again: 0.82. 400 µL of culture was mixed with 10 µL BD1807 DNA for obtaining the abrB knock out. The sample was put in a new 15 mL tube in the shaking incubator at 37 °C.
At 13:30 100 µL of culture was put on an LB plus chloramphenicol plate. This plate was put in the incubator at 37 °C. The rest of the culture was stored in the fridge at 4 °C.
11:15, 28 July 2015 - 00:00, 28 July 2015
The transformation failed, as there were no colonies on the plate.