GSM5543477: CD-tRF CT4 (scATAC-seq); Mus musculus; ATAC-seq
Mouse islets were freshly isolated at CT4 and CT16 (n=2 mice per CT and group), >100 islets were counted, and nuclei isolation was performed as noted above using ice-cold 1X lysis buffer from 10X Genomics (Pleasanton, CA). A portion of isolated nuclei were stained with Trypan Blue for manual counting and to ensure a single-nuclei suspension prior to library preparation. Approximately 5,000 nuclei per sample were subsequently aliquoted for single cell ATAC-seq (scATAC-seq) library preparation using the Chromium Next GEM Single Cell ATAC Kit (10X Genomics). 10μL of 1X Tn5 Transposition enzyme was mixed with 5uL of the diluted nuclei suspension and was incubated 50°C for 1h. The tagmented nuclei were partitioned into Gel Bead-In Emulsions (GEMs) and barcoded by unique molecular identifiers (UMIs) using the Chromium Next GEM  Single Cell ATAC library & Gel Bead kit and the C1 Chromium Instrument (10X Genomics). scATAC-seq libraries were then prepared and indexed using the Chromium i7 Sample Index kit (10X Genomics) with library size determined by Fragment Analysis (Agilent) prior to sequencing. Paired-end, dual-indexed sequencing of the libraries (50bp read #1, 8bp i7 index, 16bp i5 index, 50bp read#2) was performed using an Illumina HiSeq instrument. scATAC-seq