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DNA Shuffling

Shuffling Smear Primerless PCR

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#1 Rosario

Rosario

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Posted 10 February 2012 - 01:30 AM

Hi,

I'm working since last month with a DNA shuffling strategy to improve the enzimatic activity of 1 target enzyme.

I follow the original protocol developed from Stemmer in 1994, but I have many problems after Primerless PCR.

I always get an amazing smear of big size. I use different dilutions of this product to performe the second PCR but I've never got the right size band (2.300 kb).

Does anybody have experience with this technique? May you give me some advice?

Thanks,
RV





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