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Best way to isolate viral mRNA for RT-PCR? - (Feb/25/2013 )

I am isolating Sendai virus mRNAs, from LLC-MK2 cells, and then quantifying using RT-PCR. I have used Roche's mRNA isolation kit (using magnetic beads); however I do not find the kit to be useful, as I run out the beads fairly quickly (and i'm not very trustful of reusing previously used beads). I was wondering if there are any suggestions for: Best mRNA isolation kits? (getting a lot of use out of it)

Thanks!

-ACN-

I have had excellent results with Ribozol. Cheap and efficient.

-jerryshelly1-

Thanks for the info! Will this work for specifically isolating mRNAs and not total RNA ?

-ACN-

No total RNA. Why do you need to isolate only mRNA? If you are doing RT-PCR, it will not matter what RNA is isolated. It will only rely on your GOI from your RT cDNA. You must know the viral sequence of your gene to perform RT-PCR. Are the viral primers you are using to similar to your LLC cells? Maybe I am missing something. If you absolutely need only mRNA, try the ambion mRNA pure kit from invitrogen.

-jerryshelly1-

Yeah, you're making a lot of sense! I all ready have the specific viral primers, used for the RT-PCR, but I am not sure if they are too close to the host cells. I actually never questioned why I can't just use total RNA, I'll bring it up to my P.I. tomorrow for sure.
Thanks for the help!

-ACN-