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

  • Anna S. Dolgova
  • Marina V. Safonova
  • Vladimir G. Dedkov
Part of the Methods in Molecular Biology book series (MIMB, volume 2063)


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.

Key words

Efficient sequencing protocols dsRNA viruses Enzyme treatment Precipitation by LiCl FL amplification 


  1. 1.
    Taxonomy V (2017) The classification and nomenclature of viruses. The online (10th) report of the ICTV. Arch Virol 51(1–2):141–149Google Scholar
  2. 2.
    Aly M, Al Khairy A, Al Johani S, Balkhy H (2015) Unusual rotavirus genotypes among children with acute diarrhea in Saudi Arabia. BMC Infect Dis 15(1):192. Scholar
  3. 3.
    Shi J, Hu Z, Deng F, Shen S (2018) Tick-borne viruses. Virol Sin 33(1):21–43. Scholar
  4. 4.
    Diarra M, Fall M, Fall AG, Diop A, Lancelot R, Seck MT, .Guis H. (2018). Spatial distribution modelling of Culicoides (Diptera: Ceratopogonidae) biting midges, potential vectors of African horse sickness and bluetongue viruses in Senegal. Parasit Vectors, 11(1):341. doi: Scholar
  5. 5.
    Bournez L, Cavalerie L, Sailleau C, Breard E, Zanella G., de Almeida RS, .Hendrikx P. (2018). Estimation of French cattle herd immunity against bluetongue serotype 8 at the time of its re-emergence in 2015. BMC Vet Res, 14(1):65. doi: Scholar
  6. 6.
    Maan S, Rao S, Maan NS, Anthony SJ, Attoui H, Samuel AR, Mertens PPC (2007) Rapid cDNA synthesis and sequencing techniques for the genetic study of bluetongue and other dsRNA viruses. J Virol Methods 143(2):132–139. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Anna S. Dolgova
    • 1
  • Marina V. Safonova
    • 2
  • Vladimir G. Dedkov
    • 1
    • 3
  1. 1.Saint-Petersburg Pasteur Institute, CenterFederal Service on Consumers’ Rights Protection and Human Well-Being SurveillanceSaint-PetersburgRussia
  2. 2.Plague Control CenterFederal Service on Consumers’ Rights Protection and Human Well-Being SurveillanceMoscowRussia
  3. 3.Martsinovsky Institute of Medical Parasitology, Tropical and Vector Borne DiseasesSechenov First Moscow State Medical UniversityMoscowRussia

Personalised recommendations