Protocol for next-generation sequencing of the LSD virus genome using an amplicon-based approach

使用基于扩增子的方法对 LSD 病毒基因组进行下一代测序的协议

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作者:Rahul C Bhoyar, Bani Jolly, Harie Vignesh, Lenin Bhatt, Vigneshwar Senthivel, Ravi Israni, Vinod Scaria, Sridhar Sivasubbu

Abstract

Lumpy skin disease (LSD) is a viral disease predominantly affecting cattle caused by a poxvirus belonging to the capripoxvirus genus. Here, we present a protocol for next-generation sequencing of the LSD virus genome using an amplicon-based approach. We describe steps for DNA extraction, viral DNA enrichment, amplicon pooling and purification, and library preparation and pooling. We then detail procedures for sequencing and computational analysis. This protocol can be adapted to any Illumina sequencing platform as an accelerated and scalable system. For complete details on the use and execution of this protocol, please refer to Bhatt et al.1,2.

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