GSM9038474: RNAseq, intestinal stem cells, control-Rep3; Drosophila melanogaster; RNA-Seq
For RNAseq, after collecting midgut epithelial cells by FACS, we used a series of RNA isolation and amplification kits to satisfy the mount of mRNA for RNA-seq. RNA isolation was performed according to manufacturer's instruction (Arcturus PicoPure TM RNA isolation kit, Thermofisher, 12204-1). For ATACseq, midgut epithelial cells were collected by FACS. After centrifugation, the pelleted cells were treated by ATAC lysis buffer and centrifuged again to isolate nucleus. Then the tranposase Tn5 was used to cut opened regions on the chromosome and add a defined adaptor to these DNA fragments. For RNAseq, synthesis of cDNA complementary strands, double-stranded DNA purification, in vitro transcription of DNA into aRNA, and aRNA purification were carried out by Arcturus RiboAmp PLUS RNA Amplification kit (Thermofisher, RA7016), Arcturus cDNA Purification kit (Thermofisher, RA7011) and In-Vitro Transcription kit (Thermofisher, RA7009) respectively. For ATACseq, the tagmented DNA were purified and then amplified by PCR to construct library for next-generation sequencing.