title: Illumina HiSeq 3000 sequencing; Multi-omic analysis of gametogenesis reveals a novel signature at the promoters and distal enhancers of active genes
design: Multi-omic analysis of gametogenesis reveals a novel signature at the promoters and distal enhancers of active genes
library_construction_protocol: Enriched fractions of primary spermatocytes and round spermatids were obtained using 2 to 3 mice per experiment (i.e. 4 to 6 testes) by centrifugal elutriation. Around 1 million spermatocytes and round spermatids were used for ChIP-seq analyses. The chromatin was prepared by Diagenode using the iDeal ChIP-seq kit for histones (Diagenode Cat# C01010059) and then sheared using the Bioruptor Pico sonication device (Diagenode Cat# B01060001) combined with the Bioruptor Water cooler. ChIP was performed using the IP-Star Compact Automated System (Diagenode Cat# B03000002). The antibody H3K27cr was used at 1 µg per IP and each cell type was analyzed in two biological replicates. Libraries were prepared from input and immuno-purified DNA with 1ng as starting material using MicroPlex Library Preparation Kit v2 (Diagenode Cat# C05010013), then purified using Agencout® AMPPure® XP (Beckman Coulter) and quantified using Qubit™ dsDNA HS Assay Kit (Thermo Fisher Scientific, Q32854).
attributes.tag: Experimental Factor: cell type
attributes.value: round spermatid
attributes.tag: Experimental Factor: immunoprecipitate
attributes.value: anti-H3K27cr