title: GSM9101189: Adult peripheral blood, donor 1970; Homo sapiens; RNA-Seq
library_construction_protocol: Umbilical cord blood (UCB) mononuclear cells (MNCs) were isolated from the UCB of term deliveries (≥34 weeks of gestation) at Lucile Packard Children's Hospital-Stanford. UCB collections were performed through the Binns Program for Cord Blood Research with donor consent and institutional review board approval. Within 24 hours of collection, MNCs of UCB were obtained by density gradient separation of whole blood (Ficoll Paque Plus, GE Healthcare; 400g, room temperature, 30 minutes, deceleration off), followed by ammonium chloride red blood cell lysis (9:1 NH4Cl lysis buffer to cell suspension). Adult peripheral blood (PB) was collected from the Stanford Blood Center (SBC). Within 24 hours of collection, whole blood was diluted by 1:1 ratio with MACs buffer (1x PBS, EDTA, FBS). MNCs of PB were then obtained by density gradient separation (Ficoll Paque Plus, GE Healthcare; 400g, room temperature, 30 minutes, deceleration off), followed by ammonium chloride red blood cell lysis (9:1 NH4Cl lysis buffer to cell suspension). While our initial aims were to capture mononuclear cells, multinucleated cells such as neutrophils found in the MNC fraction were also included as part of the study. Libraries were prepared using Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 according to the manufacturer's protocol (10x Genomics), targeting 10,000 cells per sample. 12 cycles of cDNA amplification were done for all samples. Individual libraries were quality checked on an Agilent 4200 Tapestation 827 using D5000 screen tape. Next, the KAPA library quantification kit (#KK4923) was used for qPCR on a BioRad CFX96 RT PCR thermal cycler. Libraries were sequenced on the Novaseq S1 platform.