RNAseq of the blastocysts stage derived from oocytes transported in various ways
Oocyte transportation has been conducted worldwide where oocytes were transported in plastic tube containing medium in air. During the transportation period, nuclear maturation of oocytes is conducted. In this project, oocytes were incubated in three ways to mimic oocyte transportation:  oocytes are incubated in plastic tube containing culture medium in air, plastic tube containing antioxidative substrate (gellan gum) and culture medium in air or in a drop of culture medium in CO2 incubator. After 21 hours of incubation, oocytes were subjected to in vitro fertilization using a frozen thawed semen from Japanese Black bulls (Name: Isenomai) and culture up to the blastocyst stage (7 days after insemination). RNA in the embryos were extracted using RNAqueous (Invitrogen) and the RNA quality and concentration were examined using a Bioanalyzer (Agilent technologies, Palo Alto, CA, USA); a cDNA library was prepared using an NEBNextSingle Cell/Low Input RNA Library Prep Kit (Illumina, San Diego, CA, USA). One lane of the multiplied samples was sequenced as 100 bp reads (single read) using NextSeq1000 (Illumina).