mRNA-Seq of placentas bearing the OgtY851A mutation
mRNA-Seq of placentas bearing the OgtY851A mutation
Embryos were dissected at E12.5 post-coitum from the uteri of the females. Placentas were dissected in PBS + 0.1% BSA and the exterior decidua was peeled out. Clean placentas were bisected via transverse sectioning; one half was collected in a 1.5 ml tube containing 300 µl of 1x RNA Protection Reagent and snap-frozen. Homogenization of placentas and extraction of total RNA with Monarch Total RNA Miniprep Kit (NEB) followed by Turbo DNase treatment and final clean-up Library preparation was performed using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina, in combination with the NEBNext Poly(A) mRNA Magnetic Isolation Module for mRNA enrichment, according to the manufacturer's instructions.