PCR with supercoiled plasmid

Dom Spinella dspinella at chugaibio.com
Mon Dec 21 13:32:04 EST 1998


> Hi all,
> 
> Is there any problem if I amplify a DNA fragment using supercoild
> plasmid as template? OR I have to linearize the plasmid piror to PCR? Any
> experience on this is greatly appreciated.
> 
> Andrew

No problem at all. It's done all the time -- even from a colony
scraping.  However IMHO, amplification efficiency does suffer a bit with
supercoiled DNA and if you are doing anything quantitative with the data
(making reference standards for Q-PCR for example), I'd linearize the
template first. --Dom Spinella



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