2025-06-10
ENA first public
ERC000011
ENA-CHECKLIST
Andalusian Center for Development Biology (CABD)
INSDC center name
public
INSDC status
ERS23745091
SRA accession
E-MTAB-14873:Spermatogonia
Submitter Id
ArrayExpress
broker name
not collected
collection date
zebrafish
common name
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.
description
adult
developmental stage
wild type genotype
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Danio rerio
organism
Danio rerio
organism
testis
organism part
Spermatogonia
phenotype
Danio rerio
scientific_name
male
sex
Tübingen
strain
Spermatogonia
title