GSM7933407: NCS62; Homo sapiens; RNA-Seq
RNA was extracted from placental tissue as previously described.24 The Machery Nagel Nucleospin II kit was used to extract RNA from each sample and samples were stored at -80°C. Each sample was analyzed for quality control on the Agilent Bioanalyzer, with a minimum RNA Integrity Number (RIN) of 4.0 accepted for transcriptomic analysis. For each library, mRNA was purified from 10µg of total RNA using the DynaBeads mRNA Purification Kit (Invitrogen) and fragmented using the RNA Fragmentation Reagents (Ambion). Double-stranded cDNA was synthesized from fragmented mRNA using the Superscript Double-Stranded cDNA Synthesis Kit (Invitrogen) and Random Hexamer primers (50ng/ul, Invitrogen). cDNA libraries were prepared according to the manufacturer's protocol (TruSeq RNA Library Prep Kit v2) and sequenced on an Illumina Hiseq 2000. For each library, mRNA was purified from 10µg of total RNA using the DynaBeads mRNA Purification Kit (Invitrogen) and fragmented using the RNA Fragmentation Reagents (Ambion). Double-stranded cDNA was synthesized from fragmented mRNA using the Superscript Double-Stranded cDNA Synthesis Kit (Invitrogen) and Random Hexamer primers (50ng/ul, Invitrogen). cDNA libraries were prepared according to the manufacturer's protocol (TruSeq RNA Library Prep Kit v2) and sequenced on an Illumina Hiseq 2000.