A design principle for a single-stranded RNA genome that replicates with less double-strand formation

Usui, Kimihito; Ichihashi, Norikazu; Yomo, Tetsuya

There is a possibility that dsRNA is formed both during sample processing and replication. We performed RNA replication with three types of RNAs (MDV-1, 4. Loop GA10, and 22. Stem GAmis8) as described in 'Materials and Methods' section, to evaluate the amount of dsRNA formed during sample processing. T1 ribonuclease (Life Technologies), an ssRNA-specific ribonuclease, were added immediately after replication, according to a previous report (23) , to avoid further dsRNA formation. The amount of dsRNA was almost unchanged even after T1 ribonuclease treatment (Supplementary Figure S3) , which indicates that dsRNA formation during sample processing was negligible in our experiments.