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In vitro transcription not working

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#1 Ahrenhase



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Posted 01 June 2014 - 01:53 PM

I'm trying to perform T7 polyermase mediated IVT using ambion's mMEssage IVT kit (am1344).  I have performed this in the past quite frequently using digest plasmid; however, now I'm trying to use a ~140 bp PCR product (2pmol or 136ng) with the T7 promoter (TAATACGACTCACTATAGGG) attached to the 5' end.  My yields are very poor (<1ug) and when I run out on a gel, I don't see any bands.  Is the promoter I'm using sufficient?

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