GSM6592022: D14, Eprs+/-, replicate 2; Mus musculus; RNA-Seq
After digesting kidneys with 0.2 mg/ml liberase (Roche, Basel, Switzerland) at 37°C for 30 min, libraries were prepared using the chromium controller according to the 10× Chromium Next GEM Single Cell 3′ protocol (version 3.1; 10× Genomics, Pleasanton, CA, USA). Briefly, the cell suspensions were diluted in nuclease-free water to achieve a target count of 10,000. The cell suspension was mixed with master mix and loaded with gel beads and partitioning oil into a Chromium Next GEM chip G. The RNA transcripts were uniquely barcoded and reverse-transcribed within droplets. The cDNA molecules were pooled and subjected to an end repair process, the addition of a single A base, and ligation of the adapters. The products were purified and enriched using PCR to produce the final cDNA library. The purified libraries were quantified using qPCR according to the protocol (Sigma Aldrich) and were qualified using the Agilent 4200 TapeStation (Agilent Technologies, Santa Clara, CA, USA), and sequenced using HiSeqX (Illumina, San Diego, CA, USA) with a read length of 28 bp for read 1 (cell barcode and UMI), an 8-bp index read (sample barcode), and a read length of 91 bp for read 2 (RNA read). The Cell Ranger pipeline (version 3.1.0; 10× Genomics) was used to process the raw dataset.