Total RNA was extracted from the remaining tissue using the RNeasy Plus Mini Kit Endometrial tissue samples from the pregnant uteri of baboon (x3), hamster (x3), bat (x2), and squirrel (x2) were dissected and mailed to the University of Chicago in RNA-Later. These samples were further dissected to remove myometrium, luminal epithelium, and extra-embryonic tissues, and then washed 3x in ice cold PBS to remove unattached cell debris and red blood cells. Total RNA was extracted from the remaining tissue using the RNeasy Plus Mini Kit (74134, QIAGEN) per manufacturer's instructions. RNA concentrations were determined by Nanodrop 2000 (Thermo Scientific). A total amount of 2.5μg of total RNA per sample was submitted to the University of Chicago Genomics Facility for Illumina Next Gen RNA sequencing. Quality was assessed with the Bioanalyzer 2100 (Agilent). A total RNA library was generated using the TruSEQ stranded mRNA with RiboZero depletion (Illumina) for each sample. The samples were fitted with one of six different adapters with a different 6-base barcode for multiplexing. Completed libraries were run on an Illumina HiSEQ2500 with v4 chemistry on 2 replicate lanes for hamster and 1 lane for other species, of an 8 lane flow cell, generating 30-50 million 50bp single-end reads per sample.
GEO Accession
GSM4696518