Primer combo mixes

Isolated murine follicles were treated as above with 100 ng/mL FSH glycoforms and then snap-frozen in groups of 50. Total RNA was isolated from 100 snap-frozen mouse ovarian follicles treated with 100 ng/mL FSH glycoforms using RNEasy micro columns (Qiagen), and the RNA was quantified using a NanoDrop spectrophotometer (ThermoScientific) at 260 nm. Total RNA was reverse transcribed by the oligo dT method using the SuperScript III kit (ThermoScientific) as described (Wang et al., 2014 (link), Wang et al., 2015 (link), Wang et al., 2016a (link)). Taqman real time qPCR assays were performed on the cDNA samples in triplicate using custom-made or pre-inventoried primer/combo mixes (ThermoScientific or IDT). Expression of Ppil1 was used as an internal control and the relative amounts of mRNA expression were calculated as described (Wang et al., 2014 (link), Wang et al., 2015 (link), Wang et al., 2016a (link)). A list of genes interrogated is provided in Supplemental Table 1.

Total RNA was isolated from mouse ovaries or testis by RNEasy micro columns (Qiagen) and RNA quantified using a NanoDrop spectrophotometer (ThermoScientific) set at 260 nm. Approximately 500 ng-1 μg total RNA was reverse transcribed by the oligo dT method using the SuperScript III kit (ThermoScientific) as described [24 (link)-26 ]. Taqman real time qPCR assays were performed on the cDNA samples in triplicate using custom-made or pre-inventoried primer/combo mixes (ThermoScientific or IDT). Expression of Ppil1 was used as an internal control and the relative amounts of mRNA expression were calculated as described [24 (link)-26 ].