GSM7016773: Donor 1; Homo sapiens; RNA-Seq
For scRNA-seq libraries, we sorted the CD45- GYPA- BM cellular subsets from 5 patients (TF166-256-261-263-273) on a BD FACS Aria Fusion cell sorter into 384-well plates. Total RNA was extracted from samples by using the RNeasy Plus Micro Kit (Life Technologies). The libraries were generated by using the Chromium Controller Single-Cell Instrument and Chromium Single Cell 3′ Library and Gel Bead Kit v2 A Chip Kit and i7 Multiplex Kit (10x Genomics) according to the manufacturer's protocol (manual part no. CG00052 Rev A). Libraries were run on an Illumina HiSeq 8000 as 150-bp reads, at one full lane per sample. Sequencing results were demultiplexed and converted to a FASTQ format by using Illumina bcl2fastq software. Alevin was used to align to the GRCh38 transcriptome and build the (cell, UMI) expression matrix for each sample.