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RNAlater protocol

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RNA stabilization with RNAlater (Closek 05/2011, modified from Sigma and Qiagen RNAlater protocols)

1. Pour 5-10* volumes of RNAlater into an Eppie or Falcon tube (e.g. 5ml of reagent per 1mg of tissue), leaving enough room in the tube for expansion during freezing.
    *Note: 5:1 (RNAlater:tissue) is standard in most company guidelines, but Qiagen states >10 volumes of RNAlater is best.
2. Tissue should be immediately placed into the tube containing RNAlater.
    Note: The tissue should have little residual water to not dilute the reagent and be fully submerged in the RNAlater to stabalize the RNA.
3. Cap and invert/mix tube for 30seconds or until the RNAlater appears murky.
4. Place tube with sample fully submerged in RNAlater at 4C overnight*
    *Note: RNAlater should be allowed to penetrate the tissue at 4C overnight or longer. 
5. After 4C incubation, transfer tube to -20C for longterm storage

 = Tissue is stable in RNAlater for 1 month at 4C and indefinitely at -20C =