Abstract:
Next-generation sequencing (NGS) technologies are continuously being developed and are becoming a more cost-effective tool for the characterization of viral genomes. Whole genome sequencing of segmented viruses, such as epizootic hemorrhagic disease virus (EHDV), provides insights into the molecular epidemiology as well as such viral evolutionary mechanisms as genetic reassortment. Here, we present a detailed method for obtaining full genome sequence data for EHDV using Illumina technology. The protocol includes details from RNA extraction and purification, the synthesis of cDNA, sequencing library preparation, to genome assembly.
摘要:
下一代测序(NGS)技术正在不断开发,并正在成为表征病毒基因组的更具成本效益的工具。分段病毒的全基因组测序,如流行病出血性疾病病毒(EHDV),提供了对分子流行病学以及诸如基因重组等病毒进化机制的见解。这里,我们提出了使用Illumina技术获得EHDV全基因组序列数据的详细方法。该协议包括RNA提取和纯化的细节,cDNA的合成,测序文库制备,基因组组装。