GSM9572946: HT-04 Control; Homo sapiens; miRNA-Seq
Peripheral blood mononuclear cells (PBMCs) were isolated using Leucosep seperation tube. RNA was extracted using TRIzol Reagent (Invitrogen) and the SV Total RNA Isolation System (Promega, USA) following the company protocol. Small RNA libraries were prepared using the QIAseq miRNA Library Kit. The procedure consisted of the following steps: Ligation of a pre-adenylated DNA adapter to the 3′ ends and an RNA adapter to the 5′ ends of mature miRNAs. Reverse transcription using a UMI-integrated primer to tag individual molecules and introduce a universal sequence. PCR amplification using plate index kits to assign a Unique Dual Index (UDI) to each sample. Library purification using a magnetic bead-based cleanup method. Quality control assessment using an Agilent Bioanalyzer and quantification via Qubit Fluorometer. Equimolar pooling of the final libraries for sequencing on an Illumina MiSeq platform.