Next-generation sequencing of Mumps orthorubulavirus: genetic variability of mumps virus populations upon ribavirin treatment
Viral RNA was extracted from 400 L of cell culture supernatant using Quick-RNA Viral Kit (Zymo Research, USA). Isolated RNA was reversely transcribed with Superscript III (Thermo Fisher Scientific, USA) and amplified using Phusion polymerase (New England Biolabs, USA). Fragments were separated on 1% agarose gels, excised, purified using Nucleospin Gel and PCR Clean-up Kit (Macherey-Nagel, Germany) and quantified with QuantiFluor ONE dsDNA System (Promega, USA). Libraries were prepared using Illumina DNA Prep kit (Illumina, USA) following manufacturer's protocol. Libraries were quality checked on 2100 Bioanalyzer using High Sensitivity DNA Kit (Agilent, USA).