Storage of RNA

[Daniel Meier]

Peter Kristensen (ptk at biobase.aau.dk) wrote:
: Hi Netters.
: 
: I posted a similar question a few days ago, but I am not quit sure that my
: posting were send correctly, therefore I will try again.
: 
: My problem is that I am doing RT-PCR for quantitation of mRNA. The system 
: uses a standard RNA, that is made by in vitro transcription. This standard is
: added to the samples in "known" amounts.
: The problem is that I am not sure that the standard RNA is stable when stored
: at -20 C in formamide. The reason could be that it is keept in a very diluted
: form (200 pg/ul). I have felt that this would be the best way to store it since
: this allows me to thaw the same standard, thereby increasing the chance of
: adding the same amounts of standard to all samples. 
: But now it seems that I have lost my standard, that is I still add the same
: amounts to my RT-PCR but no longer getting the expected product.
: 
: Any comments and suggestions on how to store my RNA so that I can keep a 
: reliable pool of standard RNA to add.
: 
: Peter Kristensen
: Aarhus University
: Dept. of Chemistry
: Div. of Biostructural Chemistry
: 
: e-mail : ptk at biobase.aau.dk

Peter,

  Store your RNA in the appropriate concentrations in DEPC-treated water
at - 70 C.  Aliquot the RNA so that you thaw each tube once and discard. 
I wouldn't refreeze the RNA.

Good Luck,

Dan Meier
dmeier at post.its.mcw.edu