A5_1
Mus musculus
Protocols: Mouse conceptuses from control litters or blastocyst transferred litters were dissected on embryonic (E) day 10.5 of development in cold 1x phosphate buffered saline. Embryos and whole placentas were quickly phenotyped, photographed and weighed, and then snap frozen in liquid nitrogen for storage at -80C. For transferred litters, the developmental stage was consistent with that of the pseudopregnant female. Whole placentas at E10.5 were homogenized using lysing matrix D beads (MP Biomedicals). Total RNA was extracted using TRIzol reagent (Sigma-Aldrich) according to the manufacturer's instructions. Extracts were treated with DNAseI (Thermo Scientific). The concentration and purity of RNA was determined by a SpectroStar spectrophotometer (BMG LABTECH) and RNA integrity was determined by an Agilent Tapestation Bioanalyzer (Aligent Technologies LDA UK Ltd). RNA libraries were prepared by Cambridge Genomic Services (Department of Pathology, University of Cambridge) using 200 ng of total RNA and the TruSeq stranded mRNA Library Preparation kit (Illumina) according to the manufacturer's instructions. A unique index sequence was added to each RNA library to allow for multiplex sequencing.