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Able to see correct product with 2 rounds of PCR, but my PI doesn't want me


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#1 sri2010

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Posted 19 April 2011 - 07:59 PM

So after 1 round of PCR, I cannot see a product with my primers. With 2 rounds, I am able to. However, my PI doesnt want me to do this because it introduces "contamination". I'm working with human samples. (limited amounts of DNA)

Can you help me convince her?

#2 pcrman

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Posted 19 April 2011 - 08:09 PM

What PCR are you doing: MSP or BSP? For either, two rounds or nested PCR are OK. In MSP, you can first amplify the converted DNA using a pair of universal primers and then reamplify using M or U primers. For BSP, two rounds of PCR are needed for most of the times.




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