Time taken for RNA to denature - (Oct/05/2006 )

Hi everyone

Today, I leave my RNA for about 20 min at Room Temp by mistake, would this cause RNA denature or other problems to my RNA sample, and if yes, is there some other ways, that i can do to solve this problem instead of isolate new RNA

Your RNA won't denature at RT, but it could allow RNases that might have been in your sample to be active and degrade your RNA. I don't know if 20 min is enough time to do much harm though.

If you're going to just make cDNA, I would do that immedietly and not subject the RNA to any more freeze thaws and then just check the quality of the cDNA before isolating more RNA. However if the RNA is for something like Northern blotting where the quality would be very important and you don't want to waste time with degraded RNA, then I would probably prepare a new RNA sample just to be safe.

i always add RNase inhibitor as soon as my RNA is eluted, and denature at 70C for 10 mins prior to RT step