Please post: RNA in vitro transcription question
FEI YU
ad5297 at wayne.edu
Tue Jul 23 09:59:15 EST 2002
Hello, does anyone have experience to get high yielding in
vitro RNA transcription cDNA containing T7 promoter? My
problem is whenever I used NTP concentration higher than 6
mM; it completely inhibits transcription reaction. I have
tried to added polyamine and inorganic pyrohosphatase into
my reaction buffer as suggested by others; it didn't help a
lot. I know some commercial kits give very high yielding,
but they don't sell buffer separately, and all the kits are
too expensive and contains stuff I don't need, I wish to
make my own buffer to get high yielding, any thoughts?
Thanks a lot
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