GSM8478227: s2L1; Mus musculus; RNA-Seq
Mouse neonate islets were digested from pancreata and purified by Histopaque centrifugation. The isolated islets were trypsinized and sorted through flow cytometer for mCherry+ cells (β cell). The RNA extraction was performed on sorted cells via TRIzol phenol-chloroform. DNA libraries were generated using Takara's SMART-Seq v4 Low Input RNA Kit for Sequencing (Takara, Mountain View, CA) for cDNA synthesis and the Illumina NexteraXT DNA Library Preparation (Illumina, San Diego, CA) kit for cDNA dual indexing. Full-length cDNA fragments were generated from 1 to 10 ng total RNA by SMART (Switching Mechanism at 5′ End of RNA Template) technology. cDNA fragments were fragmented and dual indexed in a single step using the Nextera kit's simultaneous transposon and tagmentation step. Quality and quantity of completed libraries were assessed using Agilent DNA series chip assay (Agilent Technologies, Santa Clara, CA) and Invitrogen Qubit ds DNA HS Kit (Invitrogen, Carlsbad, CA), respectively. Each library was standardized to 2 nM.