RNA-seq mus musculus dorsal root ganglia
The amount of RNA was quantified using a Qubit 2.0 Fluorometric Quantitation system (Life Technologies) and an Experion Automated Electrophoresis System (Bio-Rad) was used to calculate RNA integrity score. RNA-seq libraries were prepared with the TruSeq Stranded mRNA LT sample preparation kit (Illumina) using Sciclone and Zephyr liquid handling robotics (PerkinElmer) at pre- and post-PCR steps. Library concentrations were determined using a Qubit 2.0 Fluorometric Quantitation system (Life Technologies) and the distribution of sizes determined using an Experion Automated Electrophoresis System (Bio-Rad). To sequence, the 14 libraries were pooled, diluted to equimolar amounts and sequenced on an Illumina HiSeq 3000/4000 using 50bp single-end chemistry. Base calls provided by the real-time analysis (RTA) software (Illumina), were converted into multiplexed, unaligned BAM format before demultiplexing into sample-specific unaligned BAM files.