Team:Evry/Notebook/Sensing/PCBs/08-18-2014
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- | <u>Sensor construction hbpR/PhbpC</u> | + | <u>Sensor construction hbpR/PhbpC:</u> |
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+ | <u>Sensor construction bphR1/bphR2:</u> | ||
+ | <br/>bphR2 gene, synthetized by Eurofins in a plasmid, was dissolved in 23,5µL of elution buffer because it was lyophilized. | ||
+ | <br/>Concentration obtained = 200ng/µL | ||
+ | <br/> | ||
+ | <br/>DH5a chimiocompetents were transformed with this plasmid according to this protocol: | ||
+ | <ol> | ||
+ | <li>Remove DH5a from -80°C (about 200µL) and let them on the ice | ||
+ | <li>Add 100ng of plasmid at DH5a and let during 30mn on ice | ||
+ | <li>Make a heat shock by putting bacteria at 42°C during 30s-1mn | ||
+ | <li>Let 1h at 37°C | ||
+ | <li>Plate 100µL on LB-agar-Amp | ||
+ | <li>Incubate at 37°C overnight, 200rpm | ||
+ | </ol> | ||
+ | |||
</p> | </p> |
Revision as of 17:50, 25 August 2014
Sensor construction hbpR/PhbpC:
Sensor construction bphR1/bphR2:bphR2 gene, synthetized by Eurofins in a plasmid, was dissolved in 23,5µL of elution buffer because it was lyophilized.
Concentration obtained = 200ng/µL
DH5a chimiocompetents were transformed with this plasmid according to this protocol:
- Remove DH5a from -80°C (about 200µL) and let them on the ice
- Add 100ng of plasmid at DH5a and let during 30mn on ice
- Make a heat shock by putting bacteria at 42°C during 30s-1mn
- Let 1h at 37°C
- Plate 100µL on LB-agar-Amp
- Incubate at 37°C overnight, 200rpm