GSM9136220: Day_5_Limbal injury, rep1; Mus musculus; RNA-Seq
Eyes were enucleated and cleaned to remove the lens and iris. The limbal region and underlying stroma, along with the adjoining conjunctival and peripheral corneal tissue (~0.5 mm), were dissected under a fluorescent binocular microscope. The collected tissue was incubated in Collagenase A (2 mg/ml) for 1 hour at 37°C, followed by a PBS wash and further incubation in Trypsin for 1 hour at 37°C. The resulting cell suspension was passed through a 40 µm cell strainer to obtain a single-cell suspension. Single-cell RNA libraries were prepared using the 10x Genomics Chromium Next GEM 3' Kit v3.1 with ~10,000 input cells per sample. Libraries were sequenced on an Illumina NextSeq 2000 (P3 100-cycle kit) with paired-end reads: 28 bp (Read 1), 90 bp (Read 2), and 10 bp indices.