title: Illumina HiSeq 2500 paired end sequencing; RNA-Seq analysis of Gestational Trophoblastic Diseases
library_construction_protocol: Sections of formalin-fixed paraffin-embedded (FFPE) tissue were freshly cut for nucleic acid extraction. RNA was extracted from FFPE tissue sections using the AllPrep DNA/RNA FFPE Kit (Qiagen, Netherlands) according to the manufacturer's protocol. RNA was eluted in 20-30 µl RNase-free water.  DNA and RNA concentrations were quantified using Qubit fluorometry (Invitrogen, Carlsbad, CA). Sequencing libraries were prepared according to manufacturer's instructions for either the TruSeq RNA Sample Prep Kit v2 or the TruSeq RNA Access Library Prep Kit (Illumina, San Diego, CA). Library concentration and size distribution were initially determined using an Agilent Bioanalyzer DNA 1000 chip (Santa Clara, CA), and Qubit fluorometry (Invitrogen, Carlsbad, CA) was performed to confirm concentration.