GSM5607886: AH2-10; Rattus norvegicus; RNA-Seq
Samples were homogenized, and RNA extracted according to the protocol, tools and reagents provided by RNeasy Mini Kit (Qiagen). Libraries were prepared according to the instructions of the NEBNext Ultra Directional RNA Library Prep kit for Illumina kit (New England Biolabs), as detailed in “Chapter 1: Protocol for use with NEBNext Poly(A) mRNA Magnetic Isolation Module”. The input yield of total RNA to start the protocol was 1 µg quantified by an Agilent 2100 Bioanalyzer using an RNA 6000 nano LabChip kit. We performed the library amplification included in the cited protocol using a PCR of 14 cycles. The obtained libraries were validated and quantified by an Agilent 2100 Bioanalyzer using a DNA7500 LabChip kit and an equimolecular pool of libraries were titrated by quantitative PCR using the “Kapa-SYBR FAST qPCR kit forLightCycler480” (Kapa BioSystems) and a reference standard for quantification.