60µl 25mM rNTP (mixed, each rNTP is 25mM)
60µl 5x transcription buffer
30µl yeast pyrophosphatase
60µl T7 polymerase
30µl 100mM DTT
20µl PCR or cut plasmid product (~300ng)
2-3 hours at 37°C
Run product on a 6%-15& denaturing gel (based on size of product)
Cut band out, elute from gel piece.
Dry pellet briefly (2 minutes)
Add store as a pellet or add low TE; either way store in the -20 in plexiglass until you need it.