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colony PCR

colony PCR - Biology Forum

colony PCR - Biology Forums. Ask questions and discuss the study of Biology. If you have Biology questions from your homework ask them here!


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  #1  
Old 11-26-2010, 02:41 PM
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Default colony PCR



Can we carry out colony PCR for low copy number vectors like pet vectors? I want to use for screening my liagted insert in the pet vector.
Also if I can use this method is it any different from regular PCR apart from introduing colony in the reaction does everything else remains the same?
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Old 12-07-2010, 09:12 AM
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Default Re: colony PCR

I regularly screen transformant colonies of pET vector derivatives using colony PCR - it works fine. The trick is to not to use too much of the colony; too much E. coli in the reaction mixture will inhibit the polymerase reaction, so less is more in this case. I normally use a 10 ul pipette tip to pick up a small amount of bacteria from the plate. Otherwise no big differences to normal PCR. Hope this helps.
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Old 12-07-2010, 12:20 PM
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Default Re: colony PCR

I knew pET was a high copy number plasmid. PCR should be ok ^^! from even a highly diluted fraction of colony lisate
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