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restriction digestion against colony PCR


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#1 mahrak

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Posted 09 May 2011 - 11:01 AM

hey every body
I wonder if any of you has experienced the situation in which I am trapped.the thing is my recombinant colonies which have shown a positive pattern( using specific primers) wouldn't give the correct fragment sizes in agarose gel electrophoresis performed right after the colony PCR.
I would be grateful for any piece of advice or hints you could suggest.

all the best
Mahsa

#2 pcrman

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Posted 09 May 2011 - 07:12 PM

Please be aware that colony PCR is not a reliable method of verifying colonies and is subject to errors such as contamination. Restriction digestion has the final say.




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