title: GSM6051736: Active; run2233; Homo sapiens; RNA-Seq
library_construction_protocol: For each sample, three biopsies from the distal esophagus of a donor were collected into RPMI medium with 10% FBS, kept on ice, and transported to the research laboratory within 30 minutes. The biopsies were then transferred into EDTA buffer (5 mM EDTA, 10% FBS, 1 mM HEPES in PBS) for 15 minutes in a 37ºC water bath, washed once with PBS, minced, and then subjected to collagenase A digestion (Roche, derived from Clostridium histolyticum, 2 mg/mL) in 10% FBS-RPMI at 37ºC for 30 minutes. The resulting suspension was diluted with 10 mL ice-cold PBS, passed through a 19-gauge needle 5 times, filtered through 2 layers of gauze, washed twice with ice-cold PBS (10 mL each), and then centrifuged at 450 g for 5 minutes, resulting in a visible, clear-edged, translucent pellet. Isolated cells were subjected to 10X Genomics sequencing in the CCHMC Gene Expression Core according to the manufacturer instructions. Bulk population cells were directly subjected to the 10X mass genomics chip (10X Genomics, Inc.), targeting 10,000 simultaneously captured live events. Each cell was uniquely barcoded during the cDNA library generation by Single Cell 3' Reagent Kits v2 per manufacturer instructions 10x scRNA-seq, Single Cell 3' v2