GSM8446775: C3; Homo sapiens; ncRNA-Seq
Blood samples were collected in 5-ml EDTA vacuum tubes, and the leukocytes were separated via red blood cell lysis solution (Solarbio, R1010). Total RNA was extracted from leukocytes with Tri®-Reagent (Sigma, T9424) within 2 hours. All procedures were performed according to the manufacturer's instructions. An RNA-seq library was constructed and sequenced by Novogene Company Limited. The NEBNext® UltraTM RNA Library Prep Kit (New England Biolabs) was used for library construction. After the library was constructed, an Agilent 2100 Bioanalyzer (Agilent Technologies) was used to determine the size of the library insert. Reverse transcription‒quantitative polymerase chain reaction (RT‒qPCR) was used to quantify the effective concentration of the library accurately to ensure quality. After qualified library inspection, Illumina sequencing was performed for the different pooled libraries according to the requirements of the effective concentration and target onboard data amount, and a 150-bp matching end reading was produced. The basic principle of sequencing is sequencing by synthesis.