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PCR reaction in cytoplasm after ODN trasfection

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#1 enri



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Posted 22 December 2010 - 07:34 AM

Hello all,
I'm working on a suspension cell line and I would like to have cytoplasmc extracts without nuclei.
I trasfected an oliginucleotide and I want to make PCR reaction on it.
Various protocols found on web site, talk about sonication and chemical or mechanical action on the cells, differential centrifugation.
Has someone a protocol with time and level of energy for sonication?
And what about chemical and mechanical...do they interfere with PCR reaction?

My goal is just to have cytoplasm without nuclei(genomic DNA).


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