Team:Groningen/Labwork/27 September 2013

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<br>The samples showed to be all positive candidates (expected product size ~2400bps).
<br>The samples showed to be all positive candidates (expected product size ~2400bps).
<br>
<br>
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<br>Colony 1 from each transformation didn't show any amylase activity and show positive results from the Colony PCR screening, therefore 100&micro;l of O/N liquid culture were inoculated in 50mL LB.
+
<br>Colony 1 from each transformation didn't show any amylase activity and show positive results from the Colony PCR screening, therefore 100 &micro;l of O/N liquid culture were inoculated in 50 mL LB.
-
<br>The cultures were induced when the OD<sub>600</sub> was 0.5 with 1mM IPTG.
+
<br>The cultures were induced when the OD<sub>600</sub> was 0.5 with 1 mM IPTG.
<br>Samples were taken every hour after induction and without any induction.
<br>Samples were taken every hour after induction and without any induction.
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<p>
<h2>Mirjam</h2>
<h2>Mirjam</h2>
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>
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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. The restriction digest went well, so gel purification can be done.
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<br><img src="https://static.igem.org/mediawiki/2013/b/bb/Pdes-CheY%2C_BBa_k823823%2C_restriction_digest_with_E-P.png" width="50%">
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<br>
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<br>Did a new transformation of CheC into the &Delta;CheY&Delta;Des knock out strain.
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<br>Started further purification of the &Delta;CheY and &Delta;CheY&Delta;Des strains on plate.
 +
</p>
</div>
</div>

Latest revision as of 19:50, 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.