IGEM:Cambridge/2008/Notebook/Bacillus/2008/08/28

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Results from Bacillus transformation with glycerol stocks

Strain added vector Antibiotic number of colonies Conclusion
IA771 none blank plate confluent colonies cells are alive
IA771 none Cm5 0 no contamination
IA771 ECE166 Cm5 2 transformation ok, very low efficiency
IA751 none blank plate confluent colonies cells are alive
IA751 none Cm5 0 no contamination
IA751 none Spc50 0 no contamination
IA751 ECE112 Cm5 0 problem, we can not manage to transform this vector
IA751 ECE153 Spc50 156 transformation ok, good efficiency


Test AmyE insertion

  • Colonies PCR for Bacillus (IA751 transformed with ECE153, IA751, and IA771)

- 20μL of lysozyme, a few colonies for each

- Cycle : 15min qt 37°C, 15min at 99°C, 1min at 4°C, 1min at 99°C and 1min at 4°C

- for each PCR : 12μL of cells, 1μL of cells, 1μL of primer1, 1μL of primer2, 5μL of master mix

- PCR (iGEM program)

- run a gel

  • Result : nothing on the gel!!! But we can not manage to PCR anything... there is a problem in our PCR protocol, or products!


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