Protocols for sequencing mumps viruses

Experimental protocols for sequencing full mumps genomes from buccal swabs

Louise Moncla1, Allison Black1,2

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

Protocols here

Manufacturer's protocols and manuals

QiAmp Viral RNA Mini Kit Handbook (Qiagen)

QiAquick Gel Extraction Kit Handbook (Qiagen)

Nextera XT DNA Library Prep Kit reference guide (Illumina)