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in vitro transcription

in vitro Transcription



Incubate for 1 hour at 37°C.

Add 10 µl STOP solution (formamide loading buffer) to each reaction, boil, and load onto a pre-run 5% denaturing polyacrylamide gel.

Run desired distance.

Cut out bands and soak in 500 µl RNA elution buffer (300 mM NaOAc, pH 6.1, 0.2% SDS, 1 mM EDTA) for 1 hour - overnight

Precipitate RNA with 2 volumes ethanol.

Resuspend in 20 µl H2O and quantitate 0.5 µl.