title: GSM8672734: MN patient NO4; Homo sapiens; RNA-Seq
library_construction_protocol: Peripheral blood were obtained and collected in heparin anticoagulant tubes. Fresh blood was separated to obtain peripheral blood mononuclear cells (PBMC) by using Histopaque-10771 (Sigma Aldrich, catalog No.10771-6 ×100 mL) according to the manufacturer's protocol. Single-cell transcriptome information was captured using the BD Rhapsody system. The single-cell suspension was randomly assigned to 200,000 micropores by a limited dilution method. The beads with oligonucleotide barcodes were added to the saturated state and paired with cells in the micropores. The cells were cleaved in micropores to hybridize mRNA molecules and the oligonucleotides on the beads were captured by bar code. After reverse transcription, ExoI digestion was performed in a test tube. During cDNA synthesis, a unique molecular identifier (UMI) was bound to each cDNA molecule at the 5' end to indicate the origin of the cell. The BD Rhapsody single-cell full transcriptome workflow includes random primer and extension (RPE), RPE amplification PCR and WTA index PCR to prepare a full transcriptome library. The high-sensitivity DNA chip (Agilent) on the bioanalyzer 2200 and the qubit high-sensitivity DNA analysis (Thermo Fisher Scientific) were used to quantify the library. The sequencing was done on a 150-bp paired-end run by an Illumina sequencer (Illumina, San Diego, CA).