Best method for freezing tissue for RT-PCR

z z-suldan at ski.mskcc.no.spam.org
Tue Jan 6 07:52:04 EST 1998


In article <34B11596.1E04 at west.bidmc.harvard.edu>,
mkoziel at WEST.BIDMC.HARVARD.EDU (Margaret Koziel MD) wrote:

> Has anyone compared different methods for freezing tissues for optimal
> preservation of mRNA (eg, snap freezing in OCT, Trizol)?
> 
> Thanks in advance,
> 
> Margaret Koziel
> Harvard Medical School
> Boston MA


I used to snap freeze my cells to -70 in residual PBS (after 1 wash) or
residual RPMI (no wash) and found inconsistent degradation. I now snap
freeze in Trizol (actually... this is what the trizol protocol says to
do... I guess I should have read the whole protocol years ago  :-) ) and
have since found very INFREQUENT degradation. I HIGHLY reccommend snap
freezing in trizol if you are not going to do the whole prep immediately.

Good luck,
Zal



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