Self-Annealing of Sendai Virus RNA

Daniel Kolakofsky, Edward Boy de la Tour, Andrée Bruschi
1974 Journal of Virology  
Both complementary strands are found in 50S Sendai virion RNA. 50S Sendai virion RNA has been shown to consist of unequal amounts of a single population of plus and minus strands by annealing studies. Preparation of [3H Jds-Sendai RNA core. [lH ]50S Sendai RNA (93.5 Mg, 1.03 x 106 counts/min, prepared as described in Fig. 1 ) was annealed in 0.75 ml of 50% formamide and 3 x buffer A (1 x buffer A: 150 mM NaCl, 20 mM Tris-Cl, pH 7.4, 1 mM EDTA) at 37 C for 90 min. The RNA was precipitated by the
more » ... addition of 2 volumes of ethanol, recovered by centrifugation, dissolved in 0.35 ml of 2x buffer A, and digested with 14 gg of ribonuclease A for 15 min at on May 7, 2020 by guest http://jvi.asm.org/ Downloaded from KOLAKOFSKY, DE LA TOUR, AND BRUSCHI 25 C. The solution was then made 0.2% in SDS, 100 ug of Proteinase K was added, and the solution was incubated for a further 10 min at 25 C. The sample was then chromatographed on a 20-ml column of Sephadex G-100 in 0.25 M LiCl, 20 mM Tris-Cl, pH 7.4, 4 mM EDTA, and 0.1% SDS, and the excluded RNA or core, representing 35% of the load, was recovered by ethanol precipitation and disolved in buffer A.
doi:10.1128/jvi.14.1.33-39.1974 fatcat:hpoxeuzyubddbjtvqha6enga6e