Team:Groningen/Labwork/27 September 2013

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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.
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.
<br><img src="https://static.igem.org/mediawiki/2013/f/f6/EstA_%2B_S3-S9.jpg" width="50%">
<br><img src="https://static.igem.org/mediawiki/2013/f/f6/EstA_%2B_S3-S9.jpg" width="50%">
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<br>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.
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Revision as of 14:37, 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 100µl of O/N liquid culture were inoculated in 50mL LB.
The cultures were induced when the OD600 was 0.5 with 1mM 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.