Transcriptome Data for dRTEL1 testes with dRTEL1-GFP

Total RNA was extracted using TRIzol reagent following the manufacturer’s instructions. 40 pairs of w1118 or dRTEL1 or dRTEL1; dRTEL1-GFP early L3 (60-72hr) stage testes were dissected in Schneider's medium (Invitrogen) and used as one set of data respectively. For RNA-Seq, each genotype was sequenced with two replicates. The integrity of RNA was confirmed by gel electrophoresis. Subsequent mRNA purification, library construction, sequencing and data analysis were performed by BGI (Beijing Genomics Institute, Shenzhen, Guangdong, China). In brief, TruSeq RNA V2/Illumina kit was used to generate the Illumina cDNA libraries. Libraries were sequenced with Illumina HiSeq 2500. Raw sequencing reads were cleaned by removing adaptor sequences, reads containing polyN sequences, and low-quality reads. The Drosophila genome (dm6, FlyBase 6.05) was used to align and filter reads.

采用TRIzol试剂（TRIzol reagent）依照厂商说明书提取总RNA。分别收集40对w1118、dRTEL1以及dRTEL1; dRTEL1-GFP基因型果蝇的早期第三龄（60-72小时）睾丸，于Schneider培养基（Schneider's medium，Invitrogen公司）中解剖，每组作为一个数据集。针对RNA测序（RNA-Seq），每个基因型设置两个生物学重复进行测序。通过凝胶电泳验证RNA的完整性。后续的mRNA纯化、文库构建、测序及数据分析均由BGI（北京基因组研究所，中国广东深圳）完成。简言之，采用TruSeq RNA V2/Illumina试剂盒（TruSeq RNA V2/Illumina kit）构建Illumina cDNA文库。使用Illumina HiSeq 2500测序平台（Illumina HiSeq 2500）对文库进行测序。通过移除接头序列、含polyN的序列以及低质量读段，对原始测序读段进行质控过滤。以果蝇基因组（dm6，FlyBase 6.05）作为参考基因组进行读段比对与过滤。