Genotyping and RT-PCR.
For genotyping, tail-clip DNA was amplified for 35 cycles. Chromosomal sex was determined by PCR with primers to the Y chromosome gene Zfy (below). The wild-type Dmrt7 allele Dmrt7+ was detected by PCR with DM5S4/DM5S5, with an annealing temperature of 60 °C. The Dmrt7flox allele was detected by PCR with DM5S5F/DM7KO7R with an annealing temperature of 62 °C. The deleted Dmrt7 allele Dmrt7− was detected with DM5S3/DM7KO7R with an annealing temperature of 62 °C. RT-PCR for Dmrt7 expression analysis was as described [23] using primers SK111/SK112 with an annealing temperature of 62 °C.