RNA-seq analysis of maternal provisioning and zygotic genome activation during early Atlantic halibut (Hippoglossus hippoglossus) embryogenesis
RNA-seq analysis of maternal provisioning and zygotic genome activation during early Atlantic halibut (Hippoglossus hippoglossus) embryogenesis
Atlantic halibut eggs were obtained from broodstock at Nordic Halibut AS (Midsund, Norway). Good-quality egg batches from four females were fertilized using cryopreserved milt and incubated in 250 L seawater incubators at 6 °C and 34 ppt salinity. Eggs were sampled at defined developmental stages (unfertilized, 8-cell, 16-cell, blastula, 25 % epiboly, 50 dd), placed in cryovials, snap frozen in liquid nitrogen, and stored at −80 °C until RNA 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 ± 0.24 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 sequencing