small RNA-seq of Homo Sapiens : EVs from Rd4 cell line
Libraries from 250ng of total RNA were prepared by using the NEBNext Multiplex Small RNA Library Prep Set 1 for Illumina (NEB, Ipswich, MA, USA) kit according to the manufacturer’s instructions. The samples were size selected on the 6% Novex TBE PAGE gel (Thermo Fisher Scientific, Waltham, MA, USA) to obtain fragments corresponding to microRNA size range. Thereafter, the products were quantified by using the KAPA Library Quantification Kit (Cat.No. KK4824, Kapa Biosystems, London, UK) and pooled at equimolar ratio. Two libraries (technical replicates) were generated in parallel, each one  containing a pool of 12 barcoded samples.The clusters were generated by using the cBot and one replicate per lane was sequenced on the HiSeq2500 (HiSeq Control Software 2.0.12.0/RTA 1.17.21.3, Illumina Inc., San Diego, CA, USA) with a 1x51 setup in RapidRun mode.