title: GSM8494442: Feline thyroid, hyperthyroid, rep7; Felis catus; RNA-Seq
library_construction_protocol: A 3-4 mm3 sample of each thyroid was dissected with scissors and washed with PBS prior to lysis in TRIzol reagent (Invitrogen). Where a distinct nodule or nodules were present in hyperthyroid cats, this area of tissue was prioritised for RNA-Seq. Phenol-chloroform extraction was used to isolate nucleic acids from the lysate, and RNA was purified from the resulting aqueous phase using a RNeasy Micro kit (Qiagen) with on-column DNase treatment to digest genomic DNA. Pure RNA was eluted with RNase-free water, and cleaned up using an RNA Clean & Concentrator-25 kit (Zymo Research). RNA integrity and concentration were determined using a Tapestation 4200 (Agilent) and Nanodrop ND-1000 (Thermo Scientific), respectively RNA-seq libraries were prepared from total RNA by Oxford Genomics Centre (Wellcome Centre for Human Genetics, University of Oxford). Following ribodepletion with the NEBNext rRNA Depletion Kit v2 (NEB) to remove abundant rRNA species, stranded libraries were generated using a TruSeq Stranded Total RNA kit (Illumina).