GSM9964652: Human periovulatory follicular aspirate, OMA (29 y), subject #197; Homo sapiens; RNA-Seq
After retrieval of cumulus-oocyte complexes, cells remaining in the follicular aspirates were pooled with cumulus cells mechanically dissociated from the oocyte. Cells underwent red blood cell lysis in ACK buffer, were resuspended, and filtered through a 40-um cell strainer to obtain a single-cell suspension. Only samples with cell viability above 85% were processed further. Single-cell cDNA libraries were generated from 0.5 x 10^6 cells using the Chromium Single Cell 3' v3 Reagent Kit (10x Genomics). Cells were partitioned into droplets and barcoded with 10x cell barcodes, UMIs, and poly(dT) sequences, then reverse-transcribed to generate barcoded cDNA and libraries per the manufacturer's protocol. Libraries were sequenced on an Illumina NovaSeq 6000 (S4 flowcell) as paired-end reads (28 cycles read 1, 8 cycles index, 150 cycles read 2).