Protocols for sequencing mumps viruses

Experimental protocols and bioinformatic pipelines for mumps genome sequencing

Louise Moncla1, Allison Black1,2, Trevor Bedford1

1Department of Epidemiology, University of Washington, Seattle, WA, USA, 2Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA

Overview of Illumina sequencing pipeline for mumps viruses

  1. vRNA extraction with QiAmp vRNA mini kit (Qiagen)
  2. cDNA synthesis with Protoscript II First strand cDNA synthesis kit (NEB)
  3. PCR with Q5 Hotstart High-fidelity DNA polymerase (NEB) and mumps-specific primers
  4. Gel extraction with QiAquick gel extraction kit (Qiagen)
  5. Library preparation with Nextera XT DNA library prep kit (Illumina)
  6. QC with Tapestation and library pooling