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Team:British Columbia/Notebook/21 July 2011
From 2011.igem.org
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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
