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RNA isolation - (Mar/25/2002 )

hi, friends,
i am doing rtpcr.  i isolate the RNA from the mesenteric artery and vein.  (it is a long time for the tissue cleaning.)   i get the RNA, add RNA inhibitor and store into the -80'C freezer.  i take it out to do one-step RT-PCR, and then store the RNA into -80 again.  i could get the result at first and cannot get it anymore soon.  the RNA is so easy to degrade? need i change to two-step to get and store the stable cDNA first?
Thanks a lot.


Repeated thawing may degrade RNA. Why don't you aliquot your tissue sample so each time just take one piece for RT-PCR?