RNA-Sequencing Analysis of the Effects of UVA Radiation on Keratinocytes

Normal human epidermal keratinocytes were treated with sham or UVA (20J/cm2) radiation in duplicate. 6h after irradiation, samples were purified using Qiagen Rneasy kit, RNA quality assessment was performed using Agilent Bioanalyzer, and library preparation was performed using oligodT selection. RNA-Seq was performed on Illumina HiSeq2000 platform with 50 bp single-end reads. Analysis used the Tuxedo pipeline. Results were validated by qRT-PCR. Overall design: Transcriptome of sham and UVA-irradiated normal human epidermal keratinocytes

将正常人表皮角质形成细胞（normal human epidermal keratinocytes）分为两组，分别进行假辐照与剂量为20J/cm²的紫外线A（UVA）辐照，每组设置两个生物学重复。辐照后6小时，使用凯杰（Qiagen）RNeasy试剂盒对样本进行总RNA纯化，采用安捷伦生物分析仪（Agilent Bioanalyzer）完成RNA质量评估，并通过寡聚dT选择法构建测序文库。随后在Illumina HiSeq2000测序平台上进行RNA测序（RNA-Seq），采用50 bp单端读长模式。转录组分析采用Tuxedo分析流程，测序结果通过实时定量聚合酶链反应（qRT-PCR）进行验证。整体实验设计：针对假辐照与UVA辐照的正常人表皮角质形成细胞开展转录组分析。