Does anyone have a good protocol for preparation of buffer for lysis of red blood cells for the extraction of RNA from whole blood??

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hi
for RNA extraction from whole blood there was a discussion at

http://www.protocol-online.org/forums/inde...?showtopic=3398
But i think that Trizol is suitable for that. Maybe you can ask Invitrogen for that.
But in nature, there is a report if RNA isolation from white blood cells (http://www.nature.com/cgi-taf/DynaPage.taf?file=/nature/journal/v429/n6987/abs/nature02451_fs.html&dynoptions=doi1109943932)


Fred

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hi Fred
im really thankfull for u,,,u answered 2 of my questions today its really nice of u
im using the qiagen extraction kit for RNA but as u know u must have a red blood cell lysis buffer in the first step ,,,i want to prepare that solutionmanually but i dont know how

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hi
i use this buffer for my cell lysis...

NaCl 150mM (3ml NaCl 1M)
EDTA pH8 2mM (160µl EDTA 250mM)
NP40 1% (2ml NP40 10%)
Tris HCl 50 mm pH 7.5
ddH2O qsp 20ml

maybe you can try it?

fred

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Hi,

The best that I tried was using RBC Lysis Solution [0.8% ammonium chloride]. Use three part of RBC for every one part of whole blood. Incubate 10 minutes at room temperature and invert once during the incubation. Centrifuge for 20 sec at maximum speed (for a volume of 300ul whole blood + 900ul RBC).

Good Luck.

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i think u should better go for enzymatic isolation using sds,edta and dnase,,,
lulu

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@ geno, NH4Cl RBC lysis solution works if the blood is fresh (unfrozen) but not if it has been frozen.

@ lula, you might want to try aquaplasmid.com's RBC Lysis Solution, and it works for both fresh and frozen blood.

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