Universal Library Preparation Protocol for Efficient High-Throughput Sequencing of Double-Stranded RNA Viruses

Methods Mol Biol. 2020:2063:181-188. doi: 10.1007/978-1-0716-0138-9_14.

Abstract

This chapter reports a library preparation protocol for efficient high-throughput sequencing of double-stranded RNA viruses. The protocol consists of four main steps, viz., enzyme treatment, precipitation using lithium chloride, full-length amplification of cDNAs, and tailing adapters for high-throughput sequencing. This protocol will be useful for all double-stranded RNA viruses and for all of the high-throughput sequencing platforms.

Keywords: Efficient sequencing protocols; Enzyme treatment; FL amplification; Precipitation by LiCl; dsRNA viruses.

MeSH terms

  • DNA Primers / genetics
  • Genome, Viral / genetics*
  • Genomic Library*
  • High-Throughput Nucleotide Sequencing / methods*
  • Nucleic Acid Amplification Techniques / methods
  • Orbivirus / genetics
  • Polymerase Chain Reaction
  • RNA Viruses / genetics*
  • RNA, Double-Stranded / genetics*
  • RNA, Viral / genetics

Substances

  • DNA Primers
  • RNA, Double-Stranded
  • RNA, Viral