GSM8894715: Detaching hematopoietic cell fraction from hTOs at D28; Homo sapiens; RNA-Seq
Cells were detached by soft mechanical dissociation from hTOs seeded with primary early thymic progenitors (ETPs) at late phase of maturation (D28) Three cell samples were incubated with TotalSeq‐B antibodies (HTO1,2,3) prior to droplet generation on the Chromium Instrument (10x Genomics). cDNA synthesis was conducted to simultaneously capture both mRNA (GEX) and HTO sequences. A targeted PCR amplification of the HTO-derived cDNA was then performed to generate a discrete HTO library, which was pooled with the GEX library for subsequent Illumina sequencing.The GEX library was prepared with Chromium Single-Cell 3′ Reagent v2 Kit (10× Genomics) as per the manufacturer's protocol. Illumina adapters were incorporated during library preparation.