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. 2015:1282:109-12.
doi: 10.1007/978-1-4939-2438-7_11.

Partial purification of IBV and subsequent isolation of viral RNA for next-generation sequencing

Sarah M Keep  1 , Erica Bickerton, Paul Britton
Affiliations

Partial purification of IBV and subsequent isolation of viral RNA for next-generation sequencing

Sarah M Keep et al. Methods Mol Biol. 2015.

Abstract

RNA viruses are known for a high mutation rate and rapid genomic evolution. As such an RNA virus population does not consist of a single genotype but is rather a collection of individual viruses with closely related genotypes-a quasispecies, which can be analyzed by next-generation sequencing (NGS). This diversity of genotypes provides a mechanism in which a virus population can evolve and adapt to a changing environment. Sample preparation is vital for successful sequencing. The following protocol describes the process of generating a high-quality RNA preparation from IBV grown in embryonated eggs and then partially purified and concentrated through a 30% sucrose cushion for NGS.

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