Northern blot with a RNA probe labeled with a fluorophore - (May/11/2018 )
I have some fluorophore labeled RNA probes that I want to try in a Northern Blot analysis using Qiagen RNeasy kit purified RNA.
I want to image the blot on the LiCor Odyssey imager/scanner. They recommend using a Northern Max kit with DIG labeling of the RNA probe prep by RT-PCR but I already have fluorescent labeled custom probes that I don't want to waste.
Every northern blot protocol I see for RNA is for DIG-labeled probes. Does any have any "protocol suggestions" for me to use these expensively made custom fluorophore RNA probes??
Basically if you have the probes already, just use those in the place of the probes in the DIG protocol. There might be some subtle differences in binding of a RNA probe vs DIG-labeled DNA, so you might have to play around with the conditions a little. If I recall correctly, there are some conditions for RNA probes somewhere in the DIG handbook.