Component	Amount per reaction	Final
Template DNA (linearized plasmid)	Variable	25 μg
10× Transcription buffer	100 μl	1×
100 mM NTP (each)*	10 μl	1 mM
10× Nucleotide analog mix	100 μl	1×
1 M DTT	10 μl	10 mM
RNase inhibitor (40 U μl−1)	10 μl	400 U
T7 RNA polymerase (home made)	50 μl	~500 U
Reaction volume		1,000 μl

^*For some analogs, the final concentration of the parent NTP should be 0.5 mM (instead of 1 mM; see Table 1).