GSM7016929: Homochronic graft (HH4-HH4), 3 hours, replicate 1; Gallus gallus; RNA-Seq
Samples were prepared for single-cell RNA sequencing as follows: tissue was dissected in cold PBS and washed thrice in cold PBS. PBS was replaced with dissociation solution (Trypsin + 0.05% EDTA, Gibco) and samples were incubated at 37ºC for 15 minutes, with trituration every 5 minutes. The dissociation reaction was terminated by addition of 10% Foetal Bovine Serum (FBS). Samples were spun down at 4ºC for 5 minutes and the supernatant discarded. Each cell pellet was resuspended in PBS + 0.25% BSA and passed through a 40μm pluriStrainer cell strainer (pluriSelect). cDNA libraries were prepared using the 10X Genomics 3'mRNA-seq workflow (10X Genomics) and sequenced using a NovaSeq instrument (Illumina).