I want to preserve breast tissues for RNA extraction later ( after 2-4 months). is it possible preserve those tissues at -80 or at -20 without preservative reagents ( such as RNA later or ALLprotect tissue Reagent )?
I freeze the tissues in liquid nitrogen just after the removal, then I keep them for several months at -80. The extractions of arn are superb!
Yes, I store mine at -80 without any reagents. But make sure they never get to thaw on the way or any time when you are doing the RNA extraction. For RNA extraction, do it in liquid nitrogen to avoid the activation of RNAses on the sample.
Thank you for all
When i can used the liquid nitrogen? do I put the tissues samples in the liquid nitrogen and then store them at -80? or store the tissues samples at -80 and liquid nitrogen used during RNA extraction.
When you are transporting your samples from the hospital/clinic make sure they are frozen. I transfer them on dry ice. Then, once in the lab, I stick them in the -80 freezer until I want to do the homogenization and extraction.
I use liquid nitrogen when doing tissue homogenization for extraction. I put liquid nitrogen to prevent it from thawing while I do the homogenization before adding lysis buffer. Never let thaw! Then once your sample is homogenized and you add the lysis buffer, the buffer prevents RNAse activity. To be safer, I add mercaptoethanol to the TRK lysis buffer as it really inhibits RNAses well.
Good luck!
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