large RNA northern - (Aug/30/2005 )
I followed ambion's protocol for microRNA isolation and thus separated small RNAs & large RNAs for different purposes. I perfromed northern on a 1.0% agarose denaturing gel and probed it with the gene of my interest as well as Ubiquitin. Did a low stringency wash but I always get probe binding with both 28S and 18S rRNA bands....IN nutshell cannot figure out between the signal for my gene......does anyone has any suggestions or comments?
Had same problem when I did northerns. Switched to a two step procedure where I extracted RNA and then used that to extract mRNA using oligio-dT cellose. Worked like a charm.
I will try it out.....