GSM9421061: C57BL/6_WT, Retina, 1; Mus musculus; RNA-Seq
Samples were obtained during surgery，the tissue were washed twice with 1× PBS and minced into approximately 1 mm3 fragments with sharp scissors and digested with collagenase Ⅱ (sigma V900892, 2 mg/mL) and DNase I (sigma DN25-100MG10 μg/mL) at 37℃ for 1h. Then, the single cell suspensions were collected through a 40 μm strainer (MACS® SmartStrainer 40μm, Miltenyi Biotec, Germany), and centrifuged at 500 g for 5 min, the supernatant was completely aspirated and cells were resuspended with 1ml 1 × PBS containing 0.04% weight/volume BSA. Finally, the cell number and viability were assessed by an automatic cell counter. Single-cell RNA-seq libraries were prepared using Single Cell 3' Library Gel Bead Kit V2 following the manufacture's introduction. Finally sequencing was performed on an Illumina Novaseq6000 with a sequencing depth of at least 100,000 reads per cell and pair end 150bp (PE150).