biostudies-arrayexpressUnknownTranscriptomicsGenomicsProteomicsNils NiepagenNanoDrop spectrophotometer, Agilent 2100 BioanalyzerAgilent 2100 Bioanalyzer (Agilent Technologies); Qubit Fluorometer (Thermo Fisher Scientific); standard electrophoresis systemnoneIllumina HiSeq 2000RNA-seq of coding RNAHippoglossus hippoglossusHippoglossus hippoglossushttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-16068RNA-seq analysis examining the effects of post-ovulatory aging on gene expression in Atlantic halibut (Hippoglossus hippoglossus) eggs. Eggs from one female were stored at 6°C for 0, 1, 2, 4, 6, or 12 hours before fertilization (t0-t12). RNA was extracted from four developmental stages: unfertilized eggs (UF), 8-cell stage (8C), blastula stage (BL), and 50 degree-days (50dd). Three technical replicates were collected for each stage and timepoint combination, resulting in 72 samples total (4 stages × 6 timepoints × 3 replicates). The study reveals that post-ovulatory aging primarily disrupts transcriptional programs at the mid-blastula transition, with significant downregulation of genes involved in axis formation, organogenesis, and developmental patterning. This dataset supports the findings that the BL stage is the critical failure window for post-ovulatory aged eggsbiostudies-arrayexpressSample Collection - Eggs were collected from a single female Atlantic halibut (Hippoglossus hippoglossus) during the reproductive spawning season at Nordic Halibut AS (Midsund, Norway). The scheduled stripping time was advanced by 12 hours compared to the commercial hatchery timing to ensure eggs were collected closer to ovulation and minimize pre-existing post-ovulatory aging. Following collection, eggs were transferred into a dry bowl, gently mixed to reduce inhomogeneity, and evenly distributed into six storage containers. Each container was filled to the top and sealed with plastic film to minimize air exposure. One fraction (t0) was fertilized immediately, while the remaining five fractions were stored at 6°C and fertilized after 1, 2, 4, 6, and 12 hours of storage (t1, t2, t4, t6, t12). Fertilization was performed using cryopreserved milt. Following fertilization, eggs were left to harden for 30 minutes, rinsed in seawater, and transferred into 800 mL cell culture flasks containing seawater with 0.25 mg/L terramycin. Incubators were maintained at 6°C in darkness. RNA samples were collected in triplicate at four developmental stages: unfertilized eggs (UF), 8-cell stage (8C, ~14 hours post-fertilization), blastula stage (BL, ~48 hours post-fertilization), and 50 degree-days (50dd, ~200 hours post-fertilization). Samples were flash-frozen in liquid nitrogen and stored for subsequent RNA extraction.Library Construction - mRNA libraries were prepared by Novogene Co. (Beijing, China) using their standard Illumina TruSeq RNA Sample Preparation protocol, following manufacturer guidelines for paired-end sequencing. Libraries were quality-checked by agarose gel electrophoresis and Bioanalyzer QC prior to sequencingNucleic Acid Extraction - Total RNA was extracted from pools of 10 eggs per sample using the RNeasy Universal Plus Mini Kit (Qiagen) following the manufacturer’s instructions. RNA concentration was measured with a NanoDrop spectrophotometer, and RNA integrity (RIN) was assessed on an Agilent 2100 Bioanalyzer, yielding mean RIN = 9.8 ± 0.24Sequencing - Sequencing was performed by Novogene Co. on an Illumina HiSeq 2500 platform using paired-end 150 bp reads. Library and run quality were assessed with Nanodrop and Agilent 2100 QC before sequencing. Data quality was verified with FastQC; average ~57 million high-quality reads per sample with > 85 % mapping rate to the Atlantic halibut reference genomeOrganizationMINSEQE ScoreAssays and DataMAGE-TAB FilesNils NiepagenfalsePost-ovulatory aging effects on Atlantic halibut (Hippoglossus hippoglossus) egg transcriptome across developmental stagesRNA-seq analysis examining the effects of post-ovulatory aging on gene expression in Atlantic halibut (Hippoglossus hippoglossus) eggs. Eggs from one female were stored at 6°C for 0, 1, 2, 4, 6, or 12 hours before fertilization (t0-t12). RNA was extracted from four developmental stages: unfertilized eggs (UF), 8-cell stage (8C), blastula stage (BL), and 50 degree-days (50dd). Three technical replicates were collected for each stage and timepoint combination, resulting in 72 samples total (4 stages × 6 timepoints × 3 replicates). The study reveals that post-ovulatory aging primarily disrupts transcriptional programs at the mid-blastula transition, with significant downregulation of genes involved in axis formation, organogenesis, and developmental patterning. This dataset supports the findings that the BL stage is the critical failure window for post-ovulatory aged eggs2026-01-30T00:00:00Z2026-01-31T02:02:04.47Z2025-11-13T15:21:48.998ZE-MTAB-16068ERP184122EFO_0002944EFO_0004170EFO_0005518EFO_0003738EFO_0004184