Protocol Online logo
Top : New Forum Archives (2009-): : Hematology

RNA isolation from whole blood - (Jan/20/2014 )

Hi all,

 

I would like to isolate RNA from whole blood.  I am planning to take blood from the donor in an EDTA tube and immediately freeze the tube in -20 deg for an hour and then in -80 deg and store until further analysis.  Do you think that this is a feasible method?

 

Thank you 

-andromeda-


AquaPreserve reagent (www.aquaplasmid.com/AquaPreserve.html) works very well for RNA extraction from frozen EDTA, ACD, heparin, and other anticogulated whole blood samples.

-AquaPlasmid-

Our RNA isolation protocol from 5-9 ml EDTA blood uses together around an hour hands-on time (lysing erytrocytes, washing, centrifuging), to transfer the washed-up leukocytes into Trizol, where they can be stored at -80 for a very long time and stable, if you have such time for that I would find it much better idea than freezing the blood (can post protocol, if interested).

 

The RNA is maybe safe while frozen to -80, but the cells are disrupted at the very moment you thaw them and nucleases will start to degrade RNA immediately. Freezing is never a good idea for RNA preservation, unless you use some kind of RNAlater or similar protective agent. Or for up to 14 days at room temperature, using the QIAGEN PAXgene tubes for the blood colection (but they are pretty expensive).

-Trof-