Partial purification of IBV and subsequent isolation of viral RNA for next-generation sequencing
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 genotypesa 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.
Keep S M, Bickerton E, Britton P
Citation: Coronaviruses: Methods and Protocols (edited by H J Maier, E Bickerton and P Britton, Humana Press) (1282), 109-112