title: Sample 1
organism: Homo sapiens
description: Protocols: Human semen samples were collected from healthy donors of unproven fertility at Nantes University Hospital (France). Total RNA was extracted using the NucleoSpin RNA XS Kit (Macherey Nagel) from 35 µL human spermatozoa cells. Quality control was realized on the RNA extracts with a 2100 bioanalyzer on chip Pico RNA (Agilent Technologies) to measure RNA Integrity Number then on nanodrop 2000 (Nanodrop) for the quantitative measurement. Reverse transcription and cDNA amplification were performed with WTA2 kit (Sigma Aldrich) from 100 ng of total RNA according to the manufacturer's instructions. Libraries were prepared from 1 ug cDNA with the NEBNext Ultra II DNA kit and Index kit Set A (Biolabs). The final pool was quantified by Qubit dsDNA High Sensitivity kit (Thermofischer) at 45 nM and qualified with the Bioanalyzer at an average size of 359 bp. To correct the unbalanced base composition of the libraries prepared, we used a PhiX spike-in (15%).