Viral transcriptome analysis of HCMV infected Kasumi-3 cells at early time post infection

Purpose: to compare viral gene expression in HCMV infected Kasumi-3 cells at 4 and 24 hours post infection Overall design: Kasumi-3 cells were infected with the TB40/E wtGFP strain of HCMV. At 4 hr post-infection, virus was inactivated and cells were either collected immediately (4hpi) or left in culture for additional 24 hours. Cells were collected and lysed in trizol. Total RNA was prepared using the Direct-zol RNA miniprep kit (Zymo Research) with an on-column DNase digestion. RNA was rRNA depleted with Ribo-zero (Illumina) and RNA-seq libraries were prepared with TrueSeq Stranded mRNA (Illumina). RNA-seq libraries were sequenced on the Illumina NextSeq 500 Sequencing System (50 bp single-end reads, 50 million reads/sample).

研究目的：比较人巨细胞病毒（HCMV）感染Kasumi-3细胞后4小时与24小时的病毒基因表达差异。
实验设计：将Kasumi-3细胞感染人巨细胞病毒TB40/E wtGFP毒株。于感染后4小时（4hpi）对病毒进行灭活处理，随后将细胞分为两组：一组即刻收集（记为4hpi组），另一组继续培养24小时后再收集。收集的细胞采用Trizol裂解，使用Direct-zol RNA迷你提取试剂盒（Zymo Research公司）并结合柱上脱氧核糖核酸酶（DNase）消化步骤制备总RNA；通过Ribo-zero（Illumina公司）去除核糖体RNA（rRNA）；采用TrueSeq链特异性mRNA建库试剂盒（Illumina公司）构建RNA测序（RNA-seq）文库。最终在Illumina NextSeq 500测序系统上完成测序，测序模式为50 bp单端读长，每个样本测序得到50 million条读段。