Vaccinia Virus Transcriptome Analysis by RNA Sequencing

RNA-sequencing (RNA-Seq) using next-generation sequencing (NGS) technique is a powerful tool for simultaneous analysis of global transcripts from both vaccinia virus and host cell. Here, we describe an RNA-Seq method for analyzing the vaccinia virus transcriptome from virus-infected HeLa cells. We pay particular attention to vaccinia virus-specific aspects of sample preparation, sequencing, and data analyses, but our method could be modified to analyze transcriptomes of other cells or tissues infected with different poxviruses.