GSM8461755: P20 Testis, Dis3cKO; Mus musculus; RNA-Seq
Single testicular cells were isolated from P20 control and knockout mice. Testes were digested into single cell suspensions with 0.25% trypsin/EDTA. scRNA-seq libraries were prepared using Chromium Single Cell 3' Reagent Kits v3 (10X Genomics) according to the manufacturer's instructions. Briefly, single cells were mixed with a suspension containing barcoded beads and UMI (unique molecular identifier) elements that allow specific tagging of messenger RNA. After partitioning thousands of cells into nanoliter-scale Gel Bead-In-EMulsions (GEMs) and barcoding, full-length barcoded cDNA was then amplified by PCR to generate sufficient mass for library construction. Libraries were constructed by fragmentation, end repair, A-tailing, adaptor ligation and index PCR. After ensuring adequate quality of the cDNA libraries, the samples were sequenced on a NovaSeq platform.