Reagent	Amount
10× Reaction Buffer	2 μL
ATP (100 mM)	2 μL
GTP (100 mM)	2 μL
CTP (100 mM)	2 μL
UTP (100 mM)	2 μL
Prepared template DNA from step 5	8 μL
T7 RNA Polymerase Mix	2 μL
Total	20 μL

Run the reaction in a thermocycler with 37°C for 8–18 h.