Molecular biology of rotaviruses. IV. Molecular cloning of the bovine rotavirus genome.

A new cloning strategy has been developed for cloning the genomes of double-stranded RNA viruses by using bovine rotavirus as a test system. The major modification adopted was the use of denatured polyadenylated double-stranded RNA as the template for reverse transcriptase. This allowed the two complementary strands of cDNA to be synthesized in a single reaction and removed the need for S1 nuclease digestion to remove the 5' hairpin structure normally generated in cDNA synthesis.