Team:Groningen/Labwork/27 September 2013

Claudio

Starch assay was performed on the 6 colonies which were inoculated yesterday (Sander).


Colony PCR was also performed on the same 6 colonies. The pair of primers used was F-hy_spank-NheI and HM14 (annealing temperature 70°C) (Sander).
The samples were checked on agarose gel 0.8% (Inne).

The samples showed to be all positive candidates (expected product size ~2400bps).

Colony 1 from each transformation didn't show any amylase activity and show positive results from the Colony PCR screening, therefore 100 µl of O/N liquid culture were inoculated in 50 mL LB.
The cultures were induced when the OD₆₀₀ was 0.5 with 1 mM IPTG.
Samples were taken every hour after induction and without any induction.

Mirjam

A colony PCR is done for EstA-S3 and EstA-S9. This showed correct clones for both of the ligation reactions. So an inoculation is done for these samples.


Did a restriction analysis on Pdes-CheY to check if the transformation is performed correctly. The same restriction digestion is done for biobrick BBa_K823823. The restriction digest went well, so gel purification can be done.


Did a new transformation of CheC into the ΔCheYΔDes knock out strain.
Started further purification of the ΔCheY and ΔCheYΔDes strains on plate.