GSM6543912: Six2+ NP from newborn Six2CreTet2/3f/f  mouse, replicate 1, WGBS; Mus musculus; Bisulfite-Seq
For scRNA-seq, kidneys were digested using Multi Tissue dissociation kit 1. Debris were removed using Miltenyi Debris Removal Solution and dead cells were removed using the Miltenyi Dead Cell Removal kit. Cell number and viability were assessed using Countess AutoCounter. For WGBS, Six2-positive cells were isolated by FACS and genomic DNA was isolated from Six2-positive cells using the DNeasy Blood & Tissue Kit. For scRNA-seq, we generated single cell suspension from whole kidney samples and loaded 10,000 cells onto the Chromium Controller (10X Genomics, PN-120223) on a Chromium Single Cell B Chip (10X Genomics, PN-120262) and processed to generate single-cell gel beads in the emulsion (GEM) according to the manufacturer's protocol (10X Genomics, CG000183). Libraries were generated using Chromium Single Cell 3' Reagent Kits v3 (10X Genomics, PN-1000092) and Chromium i7 Multiplex Kit (10X Genomics, PN-120262) according to the manufacturer's manual. Quality control for constructed library was performed by Agilent Bioanalyzer High Sensitivity DNA kit (Agilent Technologies, 5067-4626) for qualitative analysis. Quantification analysis was performed by Illumina Library Quantification Kit (KAPA Biosystems, KK4824). The library was sequenced on Illumina HiSeq 4000 system with 2x150 paired-end kits using the following read length: 28 bp Read1 for cell barcode and UMI, 8 bp I7 index for sample index and 91 bp Read2 for transcript. For WGBS, DNA concentration was measured using Picogreen. DNA quality was checked by Agilent genomic DNA screentape. The library kit is Accel-NGS® Methyl-Seq DNA Library. The library was sequenced on HiSeqX (150bp PE).