Plasma RNA Extraction and Sequencing

The RNA was extracted from 500 μL of plasma on a Maxwell 48^® RSC automat using the Maxwell^® RSC miRNA Plasma and Serum Kit (ref AS1680, Promega, Madison, WI, USA) according to the manufacturer’s protocol. Libraries for small RNA sequencing were prepared using the QIAseq miRNA Library Kit for Illumina (Qiagen, Hilden, Germany). The resulting small RNA libraries were concentrated by ethanol precipitation and quantified using a Qubit 2.0 Fluorometer (Thermo Fisher Scientific, Waltham, MA, USA) prior to sequencing on a Novaseq 6000 sequencer (Illumina, San Diego, CA, USA) with read lengths of 100 bases and 17 million single-end reads per sample, on average [24 (link),25 (link)].