RNA-seq of in vitro matured oocytes and blastocysts of buffalo

We performed RNA-seq of in vitro matured oocytes and blastocysts of buffaloes to investigate molecular aspects of in vitro maturation and embryo development. Oocytes obtained from abattoir-derived ovaries were in vitro matured and after 19 hours of incubation, the oocytes that showed the first polar body have its zona pellucida removed, and were kept in RNAlater solution until the mRNA isolation. Embryos were produced through in vitro fertilization and in vitro culture and each biological replicate was obtained from three IVP routines. Two biological replicates of pools of MII occytes (205) and pools of blastocyst (35) were sequence. The mRNA isolation was performed using Dynabeads mRNA Direct Micro Kit. Isolated mRNA were used to prepare single-end barcoded libraries with Ion Total RNA-seq Kit v2. Each library was amplified, quantified on Qubit 2.0 then sequenced in the Ion ProtonTM System (Life Technologies).