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How big a role does mixing play in PCR


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#1 Vyse

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Posted 13 August 2013 - 10:49 AM

Hello. I'd thought I'd ask whether or not mixing plays a big part in the success or failure of a PCR reactions. I tend to work with PCR mastermixes and just pipette the DNA into it then spin it down. A whirly mixer is not used.

 

Would my bands look better if I introduce a mixing step?



#2 PhalanxBio

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Posted 13 August 2013 - 10:55 AM

For PCR, I pipet up and down to mix, but this is probably not required. For qPCR in 96 well plates I never mix - I just simply add the cDNA and spin down plate.  I think the initial melting step is sufficient to mix the contents of the reaction thoroughly.  It certainly can't hurt to mix your reactions, but I'm skeptical that it will improve your bands.






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