scRNA-seq of injured cebpa mutant (CRX1816124; CRA027174)
Single-cell suspensions were prepared from freshly dissected tissue samples using enzymatic dissociation, followed by filtration to remove debris. Cell viability and concentration were assessed prior to library construction. Single-cell gene expression libraries were generated using the 10x Genomics Chromium Single Cell 3' v3.1 platform according to the manufacturer's instructions. Briefly, individual cells were partitioned into nanoliter-scale Gel Bead-In-Emulsions (GEMs), where cell lysis and reverse transcription of polyadenylated mRNA occurred. Unique cell barcodes and molecular identifiers (UMIs) were incorporated during reverse transcription. The resulting cDNA was amplified, fragmented, end-repaired, A-tailed, and adaptor-ligated. Final libraries were enriched by PCR and sequenced using paired-end reads on an Illumina NovaSeq 6000 system. The library layout was paired-end, and the platform was Illumina.