dsRNA detection in vivo
Posted 02 December 2010 - 01:18 PM
I need to detect whether my RNA of interest is present as dsRNA in vivo. I can detect both sense and antisense transcripts (more than 2kb) from the same samples using RT-PCR. I wanted to use a combination of RNases to detect the strandedness of these RNAs. Does anyone know whether Trizol is effective in maintaining the dsRNA during extraction? What about column based approaches like Mirvana?
Anyone has any experience working with DNA:RNA hybrid molecules?
Thanks in advance
Posted 02 December 2010 - 09:36 PM
sounds like you're doing in vitro detection, if it's real in vivo, you might want to use molecular beacon approach.
actually deep sequencing data revealed that most of the genome DNA are transcribed from both directions, so most RNA species have their reverse complimentary strands to go with.
Posted 03 December 2010 - 08:06 AM
Posted 05 December 2010 - 07:36 PM