title: Combining profiling of transcriptome and DNA methylome to search for candidate genes as epigenetic markers in genetically selected sea bream subjected to nutritional programming
design: Total RNA (70100 g) from liver was extracted with the MagMAXTM-96 Total RNA Isolation Kit (Applied Biosystems, Foster City, CA, United States). The RNA concentration and purity was determined using a NanoDrop 2000c Spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). Quality and integrity of the isolated RNA were checked on an Agilent Bioanalyzer 2100 total RNA Nano series II chip (Agilent, Amstelveen, Netherlands), yielding RNA integrity numbers (RINs) between 8 and 10. Samples were stored at -20 C until DNA and RNA sequencing. Illumina RNA-seq libraries were prepared from 500 ng total RNA using the Illumina TruSeq Stranded mRNA LT Sample Prep Kit (Illumina Inc. San Diego, CA, USA) according to the manufacturers instructions. All RNA-seq libraries were sequenced on an Illumina NovaSeq 6000 sequencer as 2  150 nucleotides paired-end (PE) read format according to the manufacturers protocol.