Tracking of the maternal effect gene "Filia" in bovine ovary, oocyte and invitro embryos

To find the expression of maternal effect gene Filia, two pairs of primers were designed with OLIGO5 software, one according to the whole bovine Filia mRNA sequence predicted in NCBI (access no: XM_868481.4), and the other according to its partial homology with human Filia. Oocytes were transferred to the maturation media and after 24h, matured oocytes were used for RNA extraction. Numbers of mature oocytes were inseminated and after 22h, fertilized oocytes were transferred to SOF media after cumulus cell removal. After 24h, 2-cell embryos were used for RNA extraction. Also total RNA were extracted from bovine ovaries. After cDNA synthesis, Filia and the housekeeping gene (Histone-H2a) were amplified in PCR. The reaction with first primers did not work, maybe because the predicted mRNA is not predicted in accordance with the real one. In the second reaction, Filia (164 bp) and the house keeping (240 bp) genes were amplified in the bovine ovary, oocyte and embryo. This indicates the possible role for Filia in bovine pre-implantation development.