I have conducted a test on transformation efficiency of circular plasmid DNA after some harsh storage period (in which I put the plasmid DNA in 60 Celcius degree for 2 weeks). And results showed that for a range of DNA mass for transformation (100 pg,500pg,1ng,2ng,5ng,40ng) there was no colony showed meanwhile the -20 degree stored samples showed bunch of colonies. I don't really understand the reason for this. Theoretically DNA is stable under 100 Celcius degree (as seen in PCR reaction). Could someone give me some explainations? Thank you ~
Have you performed transformation of competent cell in same time using both the plasmid (heat treated and stored at -20c)?
Yes, Gopal. For comparison purpose, it should be in same condition and same time ~
hi
long time plasmid treated in 60 c may be degrade your vector . you should before transformation, check DNA quality (Run 200ng plasmid in agarose gel 1% in two way , before and after heat treatment ) .
good luck
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