TABLE 1.
Comparison of the reaction conditions optimized in this study with those already publisheda
| Study | Buffer | Monovalent salt | Divalent salt | Act. D | Detection | [NTP] | Template |
|---|---|---|---|---|---|---|---|
| Wu and Kaesberg (26) | Tris acetate (pH 8.2) | 15 mM potassium acetate | 15 mM MgAc2 | + | [α-32P] CTP | 1 mM | Copurified/purified vRNAb |
| Wu et al. (27) | HEPES (pH 8.0) | 100 mM NaCl | 1 mM MnCl2 | − | DIG-11-UTP | 250 μM | ∼200-nt (+) sense genome fragments |
| This study | Tris acetate (pH 8.2) | 30 mM KCl | 18 mM MgCl2 | + | [α-33P]CTP | 1 mM | Copurified/purified vRNAb |
a
Act. D, actinomycin D; nt, nucleotide; NTP, nucleoside triphosphate.
b
Template RNA was either viral RNA copurified with mitochondrial fractions or viral RNA purified from virus particles or a combination of the two.