title: Sperm
organism: Danio rerio
description: Protocols: A pool of fresh testicular samples was dissected and disaggregated by a combination of mechanical and enzymatic methods. The resulting individualized cells were fixed in formaldehyde and then, sorted using a BD FACSDiscover S8 Cell Sorter. Cells were sorted according to their morphology, nucleus complexity and DNA content. Four different populations were sorted: spermatogonia, primary spermatocytes, round spermatids and sperm. Samples were dissolved in homogenization buffer (20 mM Tris pH 8.0, 100 mM NaCl, 15 mM EDTA, 1% SDS, 0.5 mg/ml Proteinase K) at 55⁰C. Two Phenol/Chlorophorm/Isoamylalcohol (25:24:1, PCI) extractions were performed. DNA was precipitated using 1/5 volume of 4 M NH4Ac and 2.5 volumes of ice-cold absolute ethanol, and 1uL of linear acrylamide. Samples were incubated overnight at -20⁰C. The DNA was pelleted and resuspended in nuclease free water. Bisulfite converted libraries were generated using the Pico Methyl-Seq Library Prep Kit (Zymo Research, Cat. D5456) following manufacturer's instructions.
attributes.tag: ENA first public
attributes.value: 2025-06-10
attributes.tag: INSDC center name
attributes.value: Andalusian Center for Development Biology (CABD)
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-14873:Sperm
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: collection date
attributes.value: not collected
attributes.tag: common name
attributes.value: zebrafish
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: testis
attributes.tag: phenotype
attributes.value: Sperm
attributes.tag: sample name
attributes.value: E-MTAB-14873:Sperm
attributes.tag: scientific_name
attributes.value: Danio rerio
attributes.tag: sex
attributes.value: male
attributes.tag: strain
attributes.value: Tübingen