Question about RACE and Exon-Exon Junction

Wenduo_Ye via methods%40net.bio.net (by yewenduo from gmail.com)
Thu Jan 19 18:28:38 EST 2012


Dear there:

I have a question I recently encountered. Particularly related to 5'RACE
and prediction of putative ATG.

The 5'RACE method I am using is RNA ligase mediated 5'RACE, in theory, only
5'Capped RNA can be identified in this method. This method was first
described on 1994.

The TSSs I identified using this method are completely different from
earlier report obtained from regular 5'RACE,I repeated my result by both
commercial(purchased from Clonetech) and homemade whole RNA. If it turns
out to be true , it will complete rewrite what people know about this gene,
so I would like to hear more about the reputation of RLM 5'RACE (I indeed
find a lot of decent papers using this methods,it seems that when RLM RACE
is engaged, northern blot confirmation is no longer needed )

Another question I am thinking quite a bit about is: anyone know if
Exon-Exon junction is allowed to present upstream of first ATG ? I Google
it but got nothing.

Thanks !!!


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