title: Illumina HiSeq 2500 sequencing; Single cell RNA-seq of adult human neural retina
design: Single cell RNA-seq of adult human neural retina
library_construction_protocol: Collection of patient samples was approved by the Human Research Ethics committee of the Royal Victorian Eye and Ear Hospital (HREC13/1151H) and Save Sight Institute (16/282) and carried out in accordance with the approved guidelines. For scRNA-seq, post-mortem eye globes were collected by the Lions Eye Donation Service (Royal Victorian Eye and Ear Hospital) for donor cornea transplantation. The remaining eye globes were used for dissection to extract the neural retina. The lens, iris and vitreous were removed and the choroid/RPE layers were excluded from the sample collection. Following extraction, the neural retinal samples were dissociated and processed for scRNA-seq right away. Neural retina samples were dissociated into single cells in dissociation solution (2mg/ml papain, 120 Units/ml DNase I) for 15 minutes. The dissociated neural retina was filtered to ensure single cell suspension using a 30µm MACS Smart Strainer (Miltenyi). For sample from Patient SC, the Dead Cell Removal kit (Miltenyi) was utilised to remove dead cells prior to scRNA-seq.  Single cells from three independent neural retina samples were captured in five batches using the 10X Chromium system (10X Genomics). The cells were partitioned into gel bead‑in‑emulsions and barcoded cDNA libraries, then prepared using the single cell 3' mRNA kit (V2; 10X Genomics).
attributes.tag: Experimental Factor: replicate
attributes.value: 1