Technique / Molecular Biology / RNA transcriptional and post-transcriptional regulation / Northern blot hybridization


Weird Northern blot hybridization



Weird Northern blot hybridization

I was doing northern analysis with arabidopsis AOS gene. When I used the
5' fragment of the gene as a probe (from the EST), it worked
well. However, if I used the 3' fragment as a probe (from PCR), it always
hybridized to all the rRNA (including 26S, 18S, 16S) but not the AOS
gene. I did ... ...

Last update 28-Jan-2002, Rating of 0 votes.

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By Nga on 28-Jan-2002
I was doing Northern hybridization using different RNA probes, I got the same specific hybridized signals of the same size (2kb) and a smear in the further down region. The mRNA are supposed to be transcribed from two separate promoters.

I guest that the mRNA is partialy degraded? I made up everything freshly in order to eliminate a chance of RNAse contamination but it does not help.

What can I do?

Thank you very much.

Nga
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