Need reliable method for RNA extraction from lymphocytes

Ben Vowels Vowels at A1.MSCF.UPENN.EDU
Wed Oct 12 16:17:17 EST 1994


Chris - 

Is it necessary to separate the lymphocytes from the rest of the node or
can you use the whole node?  If the later is possible, I would suggest
that you snap freeze the node in liq. N2 ASAP after harvesting.  Then
crush the node using a morter and pestle adding some additional liq. N2 as
necessary to keep the node from thawing.  When you have the node crushed
into a fine powder, add 1-2 ml of Trizol (Gibco) and then using an 21
gauge needle and syringe, continually aspirate and expel the tissue
solution until it is homogenous.  Then proceed with the standard Trizol
procedure.  We have used this method successfully for node RNA.  

Ben Vowels



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