IGEM:Harvard/2007/Kevin's 91r Notebook

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(New page: *Step 1: Overnight culture from Perry's frozen stock of bacteria with LppOmpA plasmid. *Step 2: Plasmid prep of LppOmpA plasmid. *Step 3: Purification of plasmid. *Step 4: Design primers ...)
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Revision as of 15:32, 26 November 2007

  • Step 1: Overnight culture from Perry's frozen stock of bacteria with LppOmpA plasmid.
  • Step 2: Plasmid prep of LppOmpA plasmid.
  • Step 3: Purification of plasmid.
  • Step 4: Design primers for PCR.
  • Step 5: Produce PCR product and run on gel.
  • Step 6: Topo cloning reaction: Mix PCR product and pTrcHis2-TOPO
  • Step 7: Transform mix into OneShot E.coli cells
  • Step 8: Overnight culture of colonies to analyze positive clones
  • Step 9: Plasmid prep of culture
  • Step 10: Digestion with Nco1 and EcoR1 and run on gel.
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