Team:British Columbia/Notebook/21 July 2011

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Team: British Columbia - 2011.igem.org

July 21 2011

(-)-limonene

  • DpnI digest yesterday's SDM PCR products at 37C for 4 hours
  • Gel verification of SDM PCR products show no bands. PCR failed.
  • PCR off the synthase to put onto yeast plasmid
    • Troubleshooting: Use gradient, from 64C to 72C
  • Gel verification of PCR products for all samples show 4 bands (same as yesterday). This may indicate, again, unspecific binding.

beta-pinene

  • Assembling biobrick part
    • Digest psb1C3 and synthase (part) with EcoRI and PstI using Biobrick Restriction Digest protocol
    • Ligate backbone and part using Biobrick Ligation protocol
    • Transform assembly into DH5alpha cells