Embryo transfer and pregnancy diagnosis
Blastocysts were stored in PBS supplemented with 20% FBS. A single
blastocyst was transferred on day 7 (estrus = day 0 = day of fusion) to
the uterine horn of each recipient animal using a non-surgical
transcervical method. Pregnancy detection was performed on day 50 post
estrus using rectal palpation and ultrasonography. Pregnant cattle were
checked by rectal palpation and ultrasonography at regular intervals
thereafter.