GSM5572087: NPY.3_962_female; Mus musculus; RNA-Seq
Skin was was placed in TRIzol reagent (Thermo Fisher Scientific, Cat# 15-596-018) and homogenized using a bead homogenizer (MP Biomedicals). Total RNA from skin was purified using the RNAEasy MiniKit (Qiagen) after phenol/chloroform separation. RNA with a RNA Integrity Number (RIN) of ≥7.0 was used for sequencing library preparation.  RNA passing quality control was converted to a sequencing ready library using the NEBNext Ultra II Directional RNA library kit as per the manufacturer's instructions (polyA mRNA workflow; NEB, Ipswich, MA). The cDNA libraries were quantitated using qPCR in a Roche LightCycler 480 with the Kapa Biosystems kit for Illumina library quantitation (Kapa Biosystems, Woburn, MA) prior to cluster generation.  Cluster generation was performed according to the manufacturers recommendations for onboard clustering (Illumina, San Diego, CA).  We generated between 30-35 million paired end 75bp sequencing reads per sample for transcript level abundance.