GSM8464831: HbSS #1 (3xNLS); Homo sapiens; RNA-Seq
Bone marrow (BM) harvested from control and treated mice after 17 weeks and purified with glycophorin-A beads to extract CD235a+ human erythroid cells. Purified cells were processed using 10X genomics according to manufacturer's protocol. CD235+ cells pooled from each mice respective to their donors and treatment. Later, cells were loaded separately on the 10x Chromium platform. Gel beads-in-emulsion (GEM) generation, cDNA amplification, and library construction were performed following the protocol of Chromium Next GEM Single Cell 3′ Reagent Kits v3.1 (Dual Index, 10x Genomics) with target recovery of 8,000–10,000 cells per library or sample. A total of 13 cycles of PCR were then conducted using the Chromium Single Cell 3′ Library and Gel Bead Kit (10x Genomics) according to the manufacturer's manuals, to amplify the cDNA library for sufficient RNA libraries. Libraries were prepared following the manufacturer's manual (10x Genomics), followed by quantification using Agilent Tapestation.