RNA isolation - (Mar/25/2002 )
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?