DNA isolation from fixed cells

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sonia's picture
DNA isolation from fixed cells

I would like to isolate DNA from ethanol fixed cells. Could anyone suggest a protocol (no kits please)

samm's picture
I just had one of our

I just had one of our undergrads try out a small expt - 80%EtOh fixed 293 cells (drop by drop ice cold EtOH, gentle continuous mixing) and unfixed ones, did the regular genomic DNA extraction from cell pellets (HotShot method), and ran a pCR. It worked just fine, so its doable. Of course, the PCR product was ~220 bp, so there might have been significant shearing (the HotShot protocol is any not really suitable for long stretches of shear sensitive gDNA). However, as a proof of principle, I'm happy to say it works!
Come to think of it, the EtOH affects membrane dynamics much more than DNA anyways.