RNA degradation

rogier stugerNOstSPAM at cellbiology.uni-frankfurt.de.invalid
Tue Jun 13 13:54:24 EST 2000


Hiya,
well kept secret: paraformaldehyde does not fix your cells. You
really need to break it up to formaldehyde before it works.
Dissolving paraformaldehyde (starting from powder) on a hot plate
is sufficient to turn it into formaldehyde.
I've used formaldehyde to fix cells, then did overnight antibody
incubations, and had no problem with the RNA I was looking for.
For a picture of the RNA, go to
http://huizen.dds.nl/~stuger/hotrnatxt.html
and download the paper (in pdf format) or go to your library (or
the plant physiology website if you have online access to the
journal) and get a copy of
Stuger et al., Messenger RNA-binding properties of nonpolysomal
ribonucleoproteins from heat-stressed tomato cells, Plant
Physiol. (1999) 120, 23-31.
(the RNA in fixed cells is on fig. 6).

Good luck,

Rogier Stuger
Mol Cell Physiol, Free U of Amsterdam
E rogier AT bio.vu.nl

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