qPCR was performed to determine the level of expression
of mRNA for ER subtype and PR in the ovaries of
each group. The relative level of expression of each mRNA
was standardized against GAPDH (internal control) as
previously described (Qiu et al., 2013). Ovaries (collected
from three goats at each group and randomly cut
two parts from each ovary) were handled as individual
samples for this analysis. Total RNA from each sample
was extracted with RNAiso Plus (Takara, DaLian, China)
following the manufacturer’s directions. Quality and quantity
of isolated total RNA were evaluated using RS323C
ultraviolet spectrophotometer (Eppendorf, Germany)