RRBS of Oryzias latipes: (OC_15) 6-month male liver
Genomic DNA was extracted using a modified column protocol. The RRBS libraries were prepared using the Diagenode Premium RRBS Kit (#C02030032, Diagenode, Denville, NJ) with the following exceptions: 200 ng of genomic DNA was used for input and the equation cycle threshold (Ct) + 2 was used to determine the number of PCR cycles needed for final library amplification. AMPure XP Beads (Beckman Coulter, Brea, CA) were used for all steps in the Diagenode protocol requiring size selection or clean-up of products. Final libraries were stored at -80C until sequencing. Libraries were submitted to the Georgia Genomics and Bioinformatics Core where they were further assessed for quality and concentration using a Fragment Analyzer (Agilent, Santa Clara, CA). Libraries were pooled and sequenced single-end for 75 cycles on one high-output flow cell of an Illumina NextSeq with 5% PhiX control (Illumina) added to provide sequence diversity.