alias: E-MTAB-3534:20090826_LUd6_15
title: Illumina Genome Analyzer IIx sequencing; iCLIP analysis of RNA bound by FUS in human brain
design: iCLIP analysis of RNA bound by FUS in human brain
library_construction_protocol: Tissue was dissociated in ice-cold PBS buffer and then subjected to UV-C irradiation. Cells were then precipitated by centrifugation for 1 min at 14000 rpm and snap-frozen on dry ice. The iCLIP protocol was followed to isolate RNA bound to FUS protein and amplify a cDNA library as described in the manuscript. The NB100-565 antibody (Novus Biologicals, US) was used at a concentration of 5 ug/mg on human brains. Specific barcodes were used for each RT reaction, with random nucleotides also incorporated into cDNAs to distinguish between PCR duplication of cDNA products; 20090826_LUd6_15: TGNNN, 20100222_LUd12_9: TGNNN, 20100222_LUd12_10: TCNNN. The final products were obtained by PCR with Illumina paired-end sequencing primers: 5-CAAGCAGAAGACGGCATACGAGATCGGTCTCGGCATTCCTGCTGAACCGCTCTTCCGATCT-3; 5-AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT-3
identifiers.namespace: DEPARTMENT OF MOLECULAR NEUROSCIENCE, UCL INSTITUTE OF NEUROLOGY, QUEEN SQUARE, LONDON WC1N 3BG, UK.  MRC LABORATORY OF MOLECULAR BIOLOGY, FRANCIS CRICK AVENUE, CAMBRIDGE CB2 0QH, UK.
identifiers.id: E-MTAB-3534:20090826_LUd6_15
identifiers.namespace: Department of Molecular Neuroscience, UCL Institute of Neurology, Queen Square, London WC1N 3BG, UK.  MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
identifiers.id: E-MTAB-3534:20090826_LUd6_15