GSM8435315: AH-F88_OFC; Rattus norvegicus; RNA-Seq
For RNA extraction, the samples were homogenized in homogenization buffer (50mM HEPES, pH 7.5, 320mM sucrose, 20mM sodium butyrate, protein and phosphatase inhibitors in DEPC water). RNA extraction was performed using a column extraction with the RNeasy Mini Kit (Qiagen) and RNase-Free DNase Set (Qiagen) for on-column DNA digestion. Libraries were prepared using the NEBNext® Poly(A) mRNA Magnetic Isolation Module and NEBNext® UltraTM II Directional RNA Library Prep for Illumina® kits (New England Biolabs), starting from 91 ng of total RNA. The instructions from "Chapter 1: Protocol for use with NEBNext® Poly(A) mRNA Magnetic Isolation Module" were followed. For library amplification as mentioned in the protocol, a 13-cycle PCR was used. For PBMCs, libraries and samples were validated and quantified using the Agilent Bioanalyzer 2100 and Agilent RNA 6000 and Agilent High Sensitivity kits. In the case of the orbitofrontal cortex, both samples and libraries were validated and quantified using the 4200 TapeStation System (Agilent) and RNA Screen Tapes and D1000 Screen Tapes kits.