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Extract RNA from fixed cells

RNA extraction

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#1 cattajulio

cattajulio

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Posted 01 November 2013 - 04:04 PM

Hi,
I fixed some primary murine  bone marrow cells with 1%PFA (10 min) and then I kept them at 4C in a solution containing 3%Newborn Calf serum, 0.02%NaAz in 1x dPBS.
I need to extract RNA from these cells for a qRT-PCR experiment.
One of my colleague has a DNA FFPE kit to extract DNA from fixed cells:I could use its Proteinase K solution.
Which buffer/ solution should I use to resuspend my fixed cells and then add Proteinase K???

I will incubate at 55C and then 80C to remove the crosslinks of the PFA. After this step  I'll use the E.Z.N.A. total RNA kit from Omega to extract RNA

Thanks
Giulio







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