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PCR reaction in cytoplasm after ODN trasfection


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#1 enri

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).

Thanks




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