Paris/July 26

From 2007.igem.org

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(Auxotrophy microscopy experiment)
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*68°C 2'
*68°C 2'
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== Auxotrophy microscopy experiment ==
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== Microscopic DAP complementation experiment ==
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We used wild type E.coli transformed by part J23107 (RFP) and ''dap-'' E.coli (w121) auxotroph to DAP. able to synthesize DAP and export it

Revision as of 14:28, 31 July 2007

Colonies PCR for cloning verification

L1.1, L1.2, L2.1, L2.2, L9.1, L10.1

L6: 8 clones

Toothpick in 50µl sterile water ==> 10'@100°C

PCR Mix:

  • 212.5 µl Master Mix
  • 8.5 µl VF2
  • 8.5 µl VR

12 µl of the mix in 13µl of lysed cells for each PCR reaction

Cycle x30:

  • 95°C 30
  • 60°C 30
  • 68°C 2'

Microscopic DAP complementation experiment

We used wild type E.coli transformed by part J23107 (RFP) and dap- E.coli (w121) auxotroph to DAP. able to synthesize DAP and export it