Gene Expression in COP1-ETV1-ETV5-cJun deficient mouse brain

Total RNA was extracted from E18.5 brains with the olfactory bulbs removed using the MirVana miRNA isolation kit (Invitrogen). RNA concentration was determined in a NanoDrop 8000 (ThermoFisher) and RNA integrity using both 2100 Bioanalyzer and 2200 TapeStation (Agilent). Libraries were prepared from 1 ug of total RNA with TruSeq RNA Sample Preparation Kit v2 (Illumina). Library size was confirmed using 2200 TapeStation and High Sensitivity D1K screen tape (Agilent) and concentration was determined by Library quantification kit (KAPA). Libraries were multiplexed five per lane and then sequenced in a HiSeq2500 (Illumina) to generate 50 million paired end 75 bp reads. The "SAMPLE_ID" sample characteristic is a sample identifier internal to Genentech. The ID of this project in Genentech's ExpressionPlot database is PRJ0013017 RNA from brain of mice with the following genotypes: COP1 het control, COP1 KO, COP1/cJun DKO, COP1/cJun/ETV1 TKO, COP1/cJun/ETV1/ETV5 QKO本研究采用MirVana miRNA分离试剂盒（MirVana miRNA isolation kit，Invitrogen），从去除嗅球的胚胎18.5天（E18.5）小鼠脑组织中提取总RNA。使用NanoDrop 8000分光光度计（NanoDrop 8000，ThermoFisher）测定RNA浓度，同时通过2100生物分析仪（2100 Bioanalyzer，Agilent）与2200 TapeStation系统（2200 TapeStation，Agilent）评估RNA完整性。取1 μg总RNA，采用TruSeq RNA样本制备试剂盒v2（TruSeq RNA Sample Preparation Kit v2，Illumina）构建测序文库；利用2200 TapeStation系统与High Sensitivity D1K筛查胶带（High Sensitivity D1K screen tape，Agilent）验证文库片段大小，并通过KAPA文库定量试剂盒（Library quantification kit，KAPA）测定文库浓度。将每泳道复用5个文库，在HiSeq2500测序仪（HiSeq2500，Illumina）上进行双端测序，生成5000万条75 bp双端读段。"SAMPLE_ID"样本特征为基因泰克（Genentech）内部使用的样本标识符；本项目在基因泰克ExpressionPlot数据库中的编号为PRJ0013017。实验所用小鼠脑组织RNA对应的基因型包括：COP1杂合对照（COP1 het control）、COP1基因敲除（COP1 KO）、COP1/cJun双敲除（COP1/cJun DKO）、COP1/cJun/ETV1三敲除（COP1/cJun/ETV1 TKO）以及COP1/cJun/ETV1/ETV5四敲除（COP1/cJun/ETV1/ETV5 QKO）。