Illumina HiSeq 2000 sequencing; CAGE-seq of human post-mortem brain tissue of the frontal lobe from patients with FTD and healthy controls
CAGE-seq of human post-mortem brain tissue of the frontal lobe from patients with FTD and healthy controls
The brain samples and/or bio samples were obtained from The Netherlands Brain Bank, Netherlands Institute for Neuroscience, Amsterdam (open access: www.brainbank.nl). All Material has been collected from donors for or from whom a written informed consent for a brain autopsy and the use of the material and clinical information for research purposes had been obtained by the NBB. Additional samples were provided by the Queen Square Brain Bank of Neurological Disorders and MRC, King College London. We complied to all relevant regulations of the mentioned brain banks for the work with these samples. Total RNA for CAGE-seq was isolated from ±100mg of frozen brain tissue with TRIzol reagent (Thermo Fischer Scientific) according to the manufacturer recommendation, followed by purification with the RNeasy mini columns (Qiagen) after DNAse treatment. CAGE-seq libraries were prepared from 5 micrograms of RNA from frozen brain tissues according to a published protocol:Takahashi, H., Lassmann, T., Murata, M. & Carninci, P. 5′ end–centered expression profiling using cap-analysis gene expression and next-generation sequencing. Nat. Protoc. 7, 542–561 (2012).