SRX382169 GSM1273824 SRP033271 GSE52665 SRS506383 GSM1273824 RNA-Seq TRANSCRIPTOMIC cDNA For improved total RNA isolation, the ejaculate was incubated at 37⁰C or immediately washed with an equal volume of Sperm Washing Medium, at 37⁰C (SWM, Irvine, Santa Ana, CA). SWM contains 21 mM HEPES buffer, 4 mM Sodium Bicarbonate, and 5 mg/mL Bovine Serum Albumin. The cell mixture was spun at 680 x g for 12 minutes. Supernatant was removed and the sperm pellet was resuspended in the residual liquid. The samples were subjected to minimal mixing and spinning in order to preserve RNA. Cell suspensions were completely homogenized by vigorous vortexing and used for RNA isolation with warmed (37⁰C) TRIzol reagent (Life Technologies, Carlsbad, CA). RNA was isolated according to the manufacturer (Invitrogen) and the RNA pellet was resuspended in RNase-free water. Total RNA was DNase treated and purified using an RNeasy Mini Kit spin column (Qiagen) and diluted in RNase-free water. Ribosomal RNA reduction was carried out via the RiboMinus Eukaryote Kit for RNA-Seq (Life Technologies). 500 ng of ribosomal-free RNA and random primers were used for whole transcriptome library preparation in total RNA-Seq protocol (Life Technologies). Final libraries were quantified using qPCR NGS Library Quantification Kit (AB SOLiD, Life Technologies). Equimolar amounts of each cDNA library were pooled and ePCR was performed using the EZBead system (Life Technologies). Enriched templated beads were applied to AB SOLiD flow cell and 75 bp sequencing reads were collected using the Solid 5500xl sequencer (Life Technologies). AB 5500xl Genetic Analyzer GEO Sample GSM1273824 http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSM1273824 gds 301273824 GEO Accession GSM1273824 SRX382170 GSM1273825 SRP033271 GSE52665 SRS506384 GSM1273825 RNA-Seq TRANSCRIPTOMIC cDNA For improved total RNA isolation, the ejaculate was incubated at 37⁰C or immediately washed with an equal volume of Sperm Washing Medium, at 37⁰C (SWM, Irvine, Santa Ana, CA). SWM contains 21 mM HEPES buffer, 4 mM Sodium Bicarbonate, and 5 mg/mL Bovine Serum Albumin. The cell mixture was spun at 680 x g for 12 minutes. Supernatant was removed and the sperm pellet was resuspended in the residual liquid. The samples were subjected to minimal mixing and spinning in order to preserve RNA. Cell suspensions were completely homogenized by vigorous vortexing and used for RNA isolation with warmed (37⁰C) TRIzol reagent (Life Technologies, Carlsbad, CA). RNA was isolated according to the manufacturer (Invitrogen) and the RNA pellet was resuspended in RNase-free water. Total RNA was DNase treated and purified using an RNeasy Mini Kit spin column (Qiagen) and diluted in RNase-free water. Ribosomal RNA reduction was carried out via the RiboMinus Eukaryote Kit for RNA-Seq (Life Technologies). 500 ng of ribosomal-free RNA and random primers were used for whole transcriptome library preparation in total RNA-Seq protocol (Life Technologies). Final libraries were quantified using qPCR NGS Library Quantification Kit (AB SOLiD, Life Technologies). Equimolar amounts of each cDNA library were pooled and ePCR was performed using the EZBead system (Life Technologies). Enriched templated beads were applied to AB SOLiD flow cell and 75 bp sequencing reads were collected using the Solid 5500xl sequencer (Life Technologies). AB 5500xl Genetic Analyzer GEO Sample GSM1273825 http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSM1273825 gds 301273825 GEO Accession GSM1273825