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in vtro transcription

Heidrun susani at sbox.tu-graz.ac.at
Tue Jan 9 04:57:58 EST 2001

I had the same problem. First I cut my plasmid with different enzymes
which produce 3'-overhang ends. Making run-off-transcription with
T7-RNA-polymerase I always got a smear from the size of my expected
fragment up to the size of the whole plasmid.
Then I tried to cut my plasmid with an enzyme which produces 5'-overhang
ends. Now my transcription produced one single band.

Poopak Banky schrieb:

> Question; i was wondering if anyone has encountered variability in RNA
> transcription products when using linearized plasmid templates?
> ---

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