Team:British Columbia/Notebook/21 July 2011

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

1,8-cineole

• Assembling biobrick part.
  • Second PCR did not work
    • Third PCR set to run overnight