Team:Evry/Notebook/Sensing/PCBs/08-18-2014

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<u>Sensor construction hbpR/PhbpC:</u>  
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<b><u>Sensor construction bphR2/PbphR1:</u></b>
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    <center>Table 1: Received primers with numbers and sequences for PCB sensing constructions </center>
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<u>Sensor construction bphR1/bphR2:</u>
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<br/>bphR2 gene, synthetized by Eurofins in a plasmid, was dissolved in 23,5µL of elution buffer because it was lyophilized.
<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/>Concentration obtained = 200ng/µL
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<li>Make a heat shock by putting bacteria at 42°C during 30s-1mn
<li>Make a heat shock by putting bacteria at 42°C during 30s-1mn
<li>Let 1h at 37°C
<li>Let 1h at 37°C
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<li>Plate 100µL on LB-agar-Amp
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<li>Plate 100µL of the culture on LB-agar-Amp
<li>Incubate at 37°C overnight, 200rpm
<li>Incubate at 37°C overnight, 200rpm
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Latest revision as of 10:53, 6 September 2014

Picture


Sensor construction bphR2/PbphR1:

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:

  1. Remove DH5a from -80°C (about 200µL) and let them on the ice
  2. Add 100ng of plasmid at DH5a and let during 30mn on ice
  3. Make a heat shock by putting bacteria at 42°C during 30s-1mn
  4. Let 1h at 37°C
  5. Plate 100µL of the culture on LB-agar-Amp
  6. Incubate at 37°C overnight, 200rpm

Aug 18

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