Illumina HiSeq 2500 sequencing; Whole genome bisulfite sequencing of bovine morula derived from metabolically divergent dairy cows
Morulas were collected at day 7 through uterine flushing. Morulas were washed and pooled in groups of 5. The samples were snap frozen and kept at -80 °C until use. morulas derived from in vitro produced 2-4 cell embryos and transferred into the oviduct of either metabolically profiled lactating cows or nulliparous heifers (as a metabolically unchallenged control) until day 7. Molulas were lysed and bisulfite treated using the EZ-DNA Methylation-Direct Kit (Zymo Research) following manufacturer's instructions.  Sequencing libraries were generated from the bisulfite treated DNA using the Pico Methyl-Seq Library Prep Kit (Zymo) according to manufacturer's recommendations with 20µM PrepAmp primer mix and 10-cycle PCR amplification.