GSM8395380: Embryonic esophagus E13.5; Mus musculus; RNA-Seq
The esophagi were microdissected at E13.5/E16.5/P0 from 12, 9, and 7 embryos/neonates, respectively, and dissected tissues pooled for each time point. Samples for each time point were isolated and pooled from 2 to 3 litters. Single- cell dissociation was done by cold active protease digestion. Dissected tissues were transferred to ice-cold PBS with 5 mM CaCl2, 10 mg/ml of Bacillus Licheniformis protease (Sigma, P5380-250MG), and 125 U/ml DNAse I (Qiagen, #79256), and incubated on ice with mixing by pipet every 2 min. Samples were incubated on ice for 7min (E13.5), 9min (E16.5) and 12min (P0) and single-cell dissociation was confirmed under a brightfield microscope. Single-cell RNA- Seq libraries for high-throughput sequencing were prepared using the Chromium Single Cell 5′ Library and Gel Bead Kit (10x Genomics).