IGEM:Cambridge/2008/Notebook/Bacillus/2008/09/04

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Result of Bacillus transformation (glycerol stock of competent cells)

strain antibiotic added vector observation conclusion
IA751 none none confluent colonies cells alive
IA751 Spc50 none nothing no contamination
IA751 Spc50 ECE153 15 ok, efficiency 25 colonies/μg of DNA
IA771 none none confluent colonies cells alive
IA771 Cm5 none nothing no contamination
IA771 Cm5 ECE166 15 ok, efficiency 5 colonies/μg of DNA

We have no contamination in our glycerol stocks. However, the efficiency of glycerol stocks seems to be lower than these of fresh competent cells.

Result of starch plates

We had 5mL of iodine solution, wait for 1min and observe.

strain vector photo observation conclusion
IA751 none
big zones of clearing for each colony ok
IA771 none
no zone of clearing for each colony ok
IA751 ECE112/none (29/08)
clearing zones for control, so no trnasformation, it is maybe E.coli, 1 clearing zone out of 3 colonies, it is certainly contamination, but that means that some colonies are transformed ok
IA751 ECE112 (02/09)
1 zone of clearing out of 4 colonies some colonies are not transformed and have a resistance to Cm5 (contamination?)
IA751 ECE153 (28/08)
no zone of clearing 5 transformed colonies
IA751 ECE153 (02/09)
2 zone of clearing out of 5 colonies some colonies are not transformed and have a resistance to Spc50 (contamination?)