In vitro transcription puzzle

Aki Hoji akhst7 at imap.pitt.edu
Thu Aug 29 11:48:07 EST 2002


Dear all, 

I made a template for generating RNA about 70bp for RNA ligase mediated
5'RACE.  I am using PCR product cut by EcoRV which generates one of PCR
products that contains T7 promoter followed by 70bp sequence.  When I threw
this template in an Ampliscribe T7 transcription kit, expecting to get 70bp
band, I got two bands, 70bp and about 140bp( or bigger) bands on the agrose
gel.  I used the gel purified EcoRV digested PCR product, but I still got
two bands.  Does anybody have any ideas as to what the hell is going on ?

Aki




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