I've been trying to extract viral RNA from tissue culture supernatant using the TRIzol LS reagent. I've followed the protocol exactly and gotten next to nothing in return. I never see a pellet and the spec reads are zilch. I re-cleaned all of my equipment and replaced all of my solutions to ensure that everything is RNase free, nothing. I tried a protocol online from UPenn that is a mixture of the TRIzol and RNeasy kit procedures, nothing. I contacted Invitrogen and they said to try diluting it in PBS to lower the serum concentration, nothing.
I know there is virus in these samples, according to plaque assay upwards of 10^9 PFU/mL. The only thing I have not tried is a carrier like glycogen. Is glycogen really going to help me that much or should I just try a product like this: https://www.roche-ap...Id=220.127.116.11.1.1
? I'd like the RNA to be usable for RT after extraction.
Any help is appreciated. Thanks.
Edited by aliquot, 27 December 2011 - 10:20 AM.