Hi ,
I would like to know the best RCLB for extraction of RNA from whole blood. It is suggested to preform bulk lysing and suspend the white cell pellet in buffer RLT before processing. I would like to know what best lysates are used. I have tried out BD FACS red cell lysing buffer however, although I was getting a good RNA quality with A260/A280 around 2.30, there was no amplification of the target gene.
If you are using the whole blood RNeasy kit... I thought it comes with the RBC lysis buffer? There shouldn't be too many differences in RBC lysing buffers. You just have to make sure that there are no residue RBCs. So your housekeeping genes didn't amplify either? Are you using targeted reverse transcription or just using random primers?
Yes, most Kits offered for whole Bllod RNA isolation contain Red Blood Cell Lysis Buffer.
If you want to buy another one, I would propose Red Blood Cell Lysis Buffer from Sigma / Roche (# 11814389001).
Sincerely
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