Need Help On Northern Blot - (Sep/26/2008 )

Hi guy,

I will be doing some result validation by confirming that the genes are expressed by nothern blotting.

RNA will extracted from a prokaryote.

After reviewing through some works, I found that RNA is harder to work with as compared with DNA.

This is my first time doing Northern Blot too, thus can anyone give me any advice on how should I go about doing, especially on the precautions to take note while handling the RNA and the RNA gel.

I would like to know some positive and negative control to be use for northern blotting too.

Thanks.

The first thing I would do is to make sure my blot detection was working, before even running the gel. Do serial dilutions of your RNA, spot it on a membrane, and practice detecting it with your probe, to establish a detection limit.