title: GSM8370246: B6_scRNASeq_1; Mus musculus; RNA-Seq
library_construction_protocol: The anterior segment of the eye including the limbus was dissected from 3-month-old Prox1-GFP mice. A strip of limbal tissue 1-2 mm wide consisting of the aqueous humor outflow tissues SC and TM, ciliary body, iris, and cornea was dissected from each anterior segment in ice-cold Dulbecco's modified Eagle's medium DMEM. To enrich for aqueous humor outflow tissues, limbal strips from 6 C57Bl/6J eyes (4 females, 2 males) were pooled for a single run of C57BL/6J scRNA-seq. C57BL/6J scRNA-seq was performed on two separate pools of eyes for a total of 12 eyes (8 females, 4 males). For 129/Sj scRNA-seq, 6 eyes (3 females, 3 males) were pooled for a single run. Single cell dissociation was performed using Papain and Deoxyribonuclease I (Worthington Biochemical Cat. no. LK003153) for 20 min at 37°C and stopped using Earl's balanced salt solution (EBSS-Thermo Fisher Scientific, Cat no. 24010-043). Cells were triturated using an 18-gauge needle, centrifuged at 300g at 4 °C, washed with cold DMEM, and filtered using a 100 μm cell strainer. Cells were resuspended in cold DMEM and placed on ice immediately. Cells were counted using Countess II automated cell counter and processed for 10x library preparation immediately