Sample handling for RNA extraction - (Jun/01/2010 )
just wanted to know if anyone evr tried RNA extraction for RT PCR in brain or other tissue samples stored at -20 degree C instead of -70. I had my mouse brain samples stored at -70 but for some space issues they got moved to -20 and reviewers are requesting some experiments including RTPCR.
it will be apain to restart new experiments and collects new brain samples.
RNA degrades at -20. You may be able to isolate some RNA from your samples and do classic RT-PCR, but it's generaly not recomended to use it for quantification experiments as the spectrum could be seriously skewed.
Even -70 isn't a best way to conserve samples for real-time quantification experiments for longer time, samples should be as soon as possible fresh frozen and stored in liquid nitrogen.