Viral RNA in total RNA eluate for transfection - (Jul/29/2014 )
Does somebody knows some methods which would allow us to bind and concentrate only viral RNA (known sequence) in total RNA eluate? We wondered if it possible to perform a direct transfection because we need an infectious virus for some experiments and we have been experiencing a lot of trouble while attempting to isolate it in usual cell culture system (virus probably dead as the sample was old and in bad condition). We are familiar with the regular laborious procedure which includes the construction of cDNA, cloning and in-vitro transcription, but is there a way to shorten it? Our virus is positive sense single-stranded RNA virus of about 7.5 kb in lenght.
If anyone has some ideas for how to do it it would be great!
Thank you in advance!
You can enrich for known sequence at the cDNA creation step by converting to cDNA using gene specific primers.