Partial purification of IBV and subsequent isolation of viral RNA for next-generation sequencing.
Partial purification of IBV and subsequent isolation of viral RNA for next-generation sequencing.
复制标题
DOI：
10.1007/978-1-4939-2438-7_11
复制
发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Britton P
中科院分区：
文献类型：
--
作者：
Keep SM;Bickerton E;Britton P
已知RNA病毒具有高突变率和快速的基因组进化。因此，RNA病毒群体不包含单一基因型，而是具有密切相关基因型的个体病毒的集合-准种，其可以通过下一代测序（NGS）进行分析。这种基因型的多样性提供了一种机制，使病毒种群能够进化并适应不断变化的环境。样品制备对于成功测序至关重要。以下方案描述了从在含胚卵中生长的IBV产生高质量RNA制备物的过程，然后通过用于NGS的30%蔗糖垫部分纯化和浓缩。
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.