Is a rat being frozen under -20 for two days still good for RNA isolation? - (Jul/28/2006 )

I want to get RNA for my qPCR. The gene I look for is abundant in the tissue.Can I still get RNA from the rat? how many RNA was degraded after two days in the freezer? Thanks

-lightgreen-

Was your rat dead when you put it in the freezer!?

Pretty nasty way to go.

-Doc_Martin-

QUOTE (Doc_Martin @ Jul 28 2006, 09:11 AM)

Was your rat dead when you put it in the freezer!?

Pretty nasty way to go.

yes, it was dead. I got the tissue I wanted and killed the rat and put it in the freezer. After two days, I realized I need to get some other tissues from the rat, and I don't have any rat that is alive.

Some people told me the RNA is still good even after one month. I doubt that. But I am not sure if it is still good after two days.

Thanks!

-lightgreen-

I'm not sure if it will be good or not, but I guess you have nothing to lose by trying to isolate RNA, especially so if you don't have any other rats.

I'd suggest that you isolate the tissue that you want without thawing the rat, then snap-freeze it in liquid nitrogen, grind the tissue with a pestle and mortar and add the tissue to your lysis solution per your standard protocol.

Just my best guess (and I don't have too much experience, so I think others here can offer you better advice) - I certaintly wouldn't thaw out the tissue though, and I'd do the isolation soon as possible, or at least get the tissue you want into -80 right away to stop the action of RNases.

Good luck!
Dave

-miRNA man-

I bet you will get it

-tertu-