Table 1.
Reagent | Volume | Control #1 | Control #2 | Template (1×) Sense | Template (1×) Antisense | Template (2×) Antisense | Template (5×) Antisense |
---|---|---|---|---|---|---|---|
10× NTP mixture | 2 μL | 2 μL | 2 μL | 2 μL | 2 μL | 4 μL | 10 μL |
10× Transcription buffer | 2 μL | 2 μL | 2 μL | 2 μL | 2 μL | 4 μL | 10 μL |
Protector RNase A Inhibitor (20 units/μL) | 1 μL | 1 μL | 1 μL | 1 μL | 1 μL | 2 μL | 5 μL |
SP6 RNA polymerase (20 units/μL), OR | 2 μL | - | 2 μL | - | - | - | - |
T7RNA polymerase (20 units/μL), OR | 2 μL | 2 μL | - | 2 μL | - | - | - |
T3 RNA polymerase (20 units/μL) | 2 μL | - | - | - | 2 μL | 4 μL | 10 μL |
Total reaction volume | 20 μL | 20 μL | 20 μL | 20 μL | 20 μL | 40 μL | 100 μL |