Team:Groningen/Labwork/27 September 2013
From 2013.igem.org
Claudio
Starch assay was performed on the 6 colonies which were inoculated yesterday (Sander).![](https://static.igem.org/mediawiki/2013/1/1d/Starch_assay_BB18_BB22.jpg)
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).
![](https://static.igem.org/mediawiki/2013/3/34/ColonyPCR_Bs_BB18_BB22_26-09-2013.jpg)
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.![](https://static.igem.org/mediawiki/2013/f/f6/EstA_%2B_S3-S9.jpg)
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.
![](https://static.igem.org/mediawiki/2013/b/bb/Pdes-CheY%2C_BBa_k823823%2C_restriction_digest_with_E-P.png)
Did a new transformation of CheC into the ΔCheYΔDes knock out strain.
Started further purification of the ΔCheY and ΔCheYΔDes strains on plate.