trypsinize cell before making RNA?

Bernhard Mayr mayr at salk.edu
Mon Dec 6 15:08:37 EST 1999


Hi,

I don't use NIH/3T3 cells, but I used Qiagen RNeasy Kit for different
cell lines. I usually took my  6-well plates straight from the incubator
to ice, sucked off the media, washed once with PBS and lysed the still
adherent cells the lysis buffer of the Qiagen RNeasy Kit (use as much
volume as will finally fit onto the column) . Then I scraped off the
lysate with a cell-scraper and followed the usual protocol. The RNA
worked fine in RT-PCR analysis.

baffulo at my-deja.com wrote:
> 
> I'm going to make RNA from NIH/3T3 cells. I wonder
> if anyone know if to trypsinize the cells before
> lysis will cause RNA degradation. According to some
> literature, even to wash cells with PBS before
> lysis will degrade RNA. It sounds a little bit
> weird to me. I'm using Qiagen RNeasy Kit.

-- 
Bernhard Mayr
The Salk Institute
Peptide Biology Labs
10010 North Torrey Pine Road
La Jolla, CA 92037
USA

Fon +1 858 453 4100 ext 1176
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