GSM4943414: CHB_Plasma_18; Homo sapiens; miRNA-Seq
Total RNA was isolated from 500μl plasma with proprietary RNA isolation protocol optimized for serum/plasma (no carrier added) (Exiqon Services, Denmark). Liver tissue (up to 5mg) was disrupted and homogenized for 4 x 1 minute at 30Hz using the TissueLyser II (Qiagen) and two 5mm stainless steel beads (Qiagen). Total RNA from liver samples was isolated using the miRNeasy Micro Kit, according to the manufacturer's protocol. A total of 5ul RNA from plasma was converted into microRNA NGS libraries using the QIAseq miRNA Library Kit (Qiagen). For liver samples, 100ng of total RNA was converted into microRNA NGS libraries. Adapters containing UMIs were ligated to the RNA and the RNA was converted to cDNA. The cDNA was amplified using PCR (22 cycles for plasma samples and 14 for liver samples) and during the PCR indices were added. The PCR samples were purified and library preparation QC was performed using either Bioanalyzer 2100 (Agilent) or TapeStation 4200 (Agilent). Based on quality of the inserts and the concentration measurements the libraries were pooled in equimolar ratios. The library pool(s) were quantified using the qPCR ExiSEQ LNA™ Quant kit (Exiqon, Denmark).
GEO Accession
GSM4943414