On 16 April 2013 00:00, DK <[email protected]> wrote: > In article <[email protected]>, Pow Joshi < > [email protected]> wrote: > >Dear All, > > > >I have some cells fixed in 4% formalin (neutral I believe). I was > wondering > >if anyone has tried any RNA extraction on such cell/tissue samples, and if > >you have any wisdom including kits, tricks, solutions, problems that you'd > >share with me. I would appreciate it hugely. > > What's the downstream application? How old are fixed samples and > how were they stored? > > I have next to zero experience with what you are facing and I am > mostly curious as to what the real answer is but my guess is that > almost all of the RNA (say, 99%) is not recoverable. >
Well, they have kits for FFPE samples that claim very high recovery. But I have never used them and didn't know the details except what's written on the kit. I would love to do some real time pcr on the samples. These cells were fixed in 2-4% formaldehyde, and kept in the refrigerator 4-8 deg C, for about 6mo to an yr. I may have some samples that were fixed for 15 min, spun down and re-suspended in PBS for storage. So the formalin exposure was minimal. yes, I too believe it may be difficult to recover any RNA from them. However, I am willing to give it a try if I have some extra information on the method(s). > Pow > > > > > _______________________________________________ > Methods mailing list > [email protected] > http://www.bio.net/biomail/listinfo/methods > _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
