I have to isolate RNA from 10 ml of -20 stored whole blood. Is it better isolate RNA from whole blood or from PBMCs?
Thank you..
The problem is that if you stored your blood at -20°C without additional manipulation, your cells will be lysed and you won't be able to isolate PBMC... So you will have to work with whole blood RNA.
Doing so I had more success using trizol than column kits (like Qiagen). But the yield remained quite low, I think the proteins coming from the erythrocytes were messing up with the extraction.
However I wouldn't advice to store your blood as it is at -20°C. separating you phase before would be better.
Hope it helps,
Good luck
Dear sir,
Phani kashyap S, Ludovic Barault thank you...
suggest me both protocols
i tried PBMC isolation using ammonium chloride method followed by trizol RNA isolation but am getting less yield 30-40 ng/ul and DNA contamination.
and with Qiagenkit the con. is too low i.e 5-7 ng/ul
Dear Govardhan,
Have a look at this:
https://www.ncbi.nlm.nih.gov/pubmed/24567882
Good luck,
Hamid
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