title: GSM4458991: B_Control_rep2; Rattus norvegicus; RNA-Seq
library_construction_protocol: RNA extraction was performed with the Qiagen RNEasy Miniprep kit according to the manufacturer protocol (Qiagen, Cat. 74134).  RNA quality was determined using an Agilent 2100 Bioanalyzer and all samples, for both tissue types, were found to have an RNA integrity number (RIN) ≥ 10.  Sequencing libraries were prepared as described previously, using NEBNext Ultra Directional RNA Library Prep kit and NEBNext Poly(A) mRNA Magnetic Isolation Module (catalogs E7420 and E7490; New England Biolabs), to be compatible with Illumina sequencing.  Isolation, heat fragmentation, and priming was performed according to manufacturer's instructions.  cDNA synthesis was followed by purification and size selection.  Finally, library clean-up was performed used AMPure XP beads (Beckman Coulter Genomics, Cat. A63881) and quality was assessed using the Agilent 2100 Bioanalyzer.  For each tissue library sequencing was performed using a 125-bp single-end protocol on a single lane of an Illumina HiSeq2500 sequencer.  Approximately, 27.4 million uniquely mapped reads were generated per placental library, and 23.7 million reads per fetal forebrain library. Sequencing libraries were prepared using NEBNext Ultra Directional RNA Library Prep kit and NEBNext Poly(A) mRNA Magnetic Isolation Module (catalogs E7420 and E7490; New England Biolabs), to be compatible with Illumina sequencing.