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isolation of an already frozen tissue sample

RNAlater -80C

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#1 myself

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Posted 08 February 2012 - 02:07 AM

Hi,

I have a set of samples from pituitary adenomas that were put in -80 fridge (without snap freezing) right after the surgery less than a week ago without any preserving solution like RNA later or anything else. Now I want to isolate my RNA with preserving maximum quality.
What to do? I have RNA later solution and TRizol in my lab?

Should I add Trizol to the frozen tissue and immediately procede with the islolation to achieve the best results or do you have any other suggestions?
Does the fact that it was done without snap freezing will considerably affect my RNA quality?

Cheers

#2 Trof

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Posted 08 February 2012 - 02:09 PM

You can try RNA-later ICE, not normal RNA-later, but I'm affraid slow freezing already started to degrade RNA. And more storing it in -80, though it may not be such difference for a week.
You can crush the frozen tissue in liquid nitrogen with mortar and pestle, and then add Trizol, that 's the best way to preserve what is left.

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