Small RNA Sequencing from Plasma

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, 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). Samples were indexed in batches of 96, with a targeted sequencing depth of 17 million reads per sample. Sequencing was performed using 100 base single-end reads, using an Novaseq6000 sequencer (Illumina, San Diego, CA, USA) [35 (link),36 (link)].