Genomic DNA was sheared into fragments ranging from 200–300 bp using a Bioruptor ultrasonicator (Diagenode, Seraing, Belgium). A custom capture array (NimbleGen SeqCap®, Roche) was designed and used to capture all coding exons, splice sites, and the flanking intron sequences of THSD4 gene (NM_024817) and 23 other genes already known to be associated with Marfan syndrome and associated disease and heritable thoracic aneurysm and dissections. Amplification, sequencing, and variant annotation were performed as previously described.8 (link),11 (link),15 (link) All variants of interest were systematically confirmed by bidirectional Sanger sequencing primers (in Supplementary table S1). Additional details concerning materials and methods are found in the Supplementary Information.
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Elbitar S., Renard M., Arnaud P., Hanna N., Jacob M.P., Guo D.C., Tsutsui K., Gross M.S., Kessler K., Tosolini L., Dattilo V., Dupont S., Jonquet J., Langeois M., Benarroch L., Aubart M., Ghaleb Y., Khalil Y.A., Varret M., Khoury P.E., Ho-Tin-Noé B., Alembik Y., Gaertner S., Isidor B., Gouya L., Milleron O., Sekiguchi K., Milewicz D., Backer J.D., Le Goff C., Michel J.B., Jondeau G., Sakai L.Y., Boileau C, & Abifadel M. (2020). Pathogenic variants in THSD4, encoding the ADAMTS-like 6 protein, predispose to inherited thoracic aortic aneurysm. Genetics in medicine : official journal of the American College of Medical Genetics, 23(1), 111-122.
Other organizations :
Ghent University Hospital, The University of Texas Health Science Center at Houston, Protein Research Foundation, Osaka University, Institute of Cancer Research, Centro San Giovanni di Dio Fatebenefratelli, Istituti di Ricovero e Cura a Carattere Scientifico, Centre de Recherche en Myologie, Hôpital Necker-Enfants Malades, Hôpitaux Universitaires de Strasbourg, Université de Strasbourg, Centre Hospitalier Universitaire de Nantes, Oregon Health & Science University, Shriners Hospitals for Children - Portland
Shearing of genomic DNA into fragments ranging from 200–300 bp using a Bioruptor ultrasonicator
dependent variables
Sequencing of the captured coding exons, splice sites, and flanking intron sequences of THSD4 gene and 23 other genes associated with Marfan syndrome and related diseases
control variables
Custom capture array (NimbleGen SeqCap®, Roche) used to capture all coding exons, splice sites, and flanking intron sequences of THSD4 gene and 23 other genes
Amplification, sequencing, and variant annotation performed as previously described in the cited references
Annotations
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