GSM6244219: Ovarian follicular aspirate, normal, rep2; Homo sapiens; RNA-Seq
Immediately after retrieval of cumulus-oocyte complexes, the cells in the aspirates were subjected to red blood cell lysis in an ACK RBC lysis buffer to remove erythrocytes. The remaining cells were thoroughly resuspended and filtered through a 40-µm cell strainer to dissociate cell clumps into single-cells. Only samples showing acceptable viability (above 85%) were subjected to the following processes. 0.5 x 10^6 cells were used to generate single-cell cDNA libraries using the Chromium Single Cell 3' v3 Reagent kit at the Art & Science Imaging Center of the University of Kentucky. Briefly, individual cells were separated into droplets and barcoded in gel beads, including 10x cell barcodes, unique molecular identifier (UMI), and poly(dT) sequences. Reverse transcription reactions were engaged to generate barcoded full-length cDNA.