GSM5931429: embryonic hindlimb bud scRNA-seq; Mus musculus; RNA-Seq
Microdissected hindlimb tissues were collected from 10 embryos at E10.5 (37-40 somites) in cold PBS. Tissues were dissociated to single cells using an enzymatic cocktail of Liberase and DNase I for 10 minutes at 37 degrees. Cells were passed through a 45 µm strainer to remove clumps and ensure a single cell suspension. Dead cells were eliminated with a 'Dead Cell Removal Kit' with magnetic beads (MACS, Milteny Biotech). A single cell suspension of pooled live hindlimb cells were loaded into one well for single-cell capture using the Chromium Single Cell 3' Reagent Kit V2 (10X Genomics). Library preparation for each sample was also performed using the Chromium Single Cell 3' Reagent Kit V2, and each sample was given a unique i7 index. Libraries were pooled and subjected to sequencing in an Illumina NovaSeq sequencer.