Re: RBC lysis when using buffy coats -> how?

From: smonard@adarc.org
Date: Mon Mar 22 1999 - 09:59:24 EST


Hi

The standard red cell lysing solution for human cells is the ammonium
chloride lysing solution. We make up a 10X stock and dilute as required.
The recipe to make 1 liter of 10X stock is as follows:

80.2g NH4Cl
8.4g NaHCO3
1mM EDTA
adjust pH to 7.4
add dH20 to 1 liter

We use it to lyse whole blood, adding about 10 times the volume of solution
as blood and incubate for about 10-15 minutes on ice. Others use it room
temp or at 37 C. I understand red cells are harder to lyse if they have
been stored at 4 degrees C. Immature red cells are also hard to lyse.

I'll package some up and call it LyseMaster or TurboLyse and sell it you
for 100bucks for 10mls if it would make you feel better!

Simon Monard
ADARC
NYC



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