Hi everyone
I am trying to run an immunoblot for dsRNA molecules of specific lengths. I have been running 5% PAGE gels and transferring the RNA using semi-dry transfer @ less than 10V. Note: the gel is a small size gel used for regular protein gels.
After transferring, I have not been able to a get a signal for the specific length of dsRNA molecule which I detect using a dsRNA specific antibody. If anyone has run a RNA immunoblot. The transfer seems to work well as I can pick up signal for my reference lane (+ve ctrl) but not for the lanes where I have loaded RNA extracts from cells.
I request you to help me with some suggestions.
We described a method for dsRNA immunoblotting in Schonborn et al. (1991)
Nucl. Acids Res. 19 (11): 2993-3000. doi: 10.1093/nar/19.11.2993
The method was used by many students since then and it works well. If you describe the exact methodology of your experiment, I might help.
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