title: Validation of Suspected Somatic Single Nucleotide Variations in the Brain of Alzheimer's Disease Patients.
abstract: Next-generation sequencing techniques and genome-wide association study analyses have provided a huge amount of data, thereby enabling the identification of DNA variations and mutations related to disease pathogenesis. New techniques and software tools have been developed to improve the accuracy and reliability of this identification. Most of these tools have been designed to discover and validate single nucleotide variants (SNVs). However, in addition to germ-line mutations, human tissues bear genomic mosaicism, which implies that somatic events are present only in low percentages of cells within a given tissue, thereby hindering the validation of these variations using standard genetic tools. Here we propose a new method to validate some of these somatic mutations. We combine a recently developed software with a method that cuts DNA by using restriction enzymes at the sites of the variation. The non-cleaved molecules, which bear the SNV, can then be amplified and sequenced using Sanger's technique. This procedure, which allows the detection of alternative alleles present in as few as 10% of cells, could be of value for the identification and validation of low frequency somatic events in a variety of tissues and diseases.
authors.affiliation: Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CIBERNED), ISCIII, Madrid, Spain.|Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Madrid, Spain.
authors.forename: Alberto
authors.initials: A
authors.lastname: Gomez-Ramos
authors.affiliation: Sygnis S.L.U. Parque Científico de Madrid. Cantoblanco, Madrid, Spain.
authors.forename: Angel J
authors.initials: AJ
authors.lastname: Picher
authors.affiliation: Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CIBERNED), ISCIII, Madrid, Spain.|Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Madrid, Spain.
authors.forename: Esther
authors.initials: E
authors.lastname: García
authors.affiliation: Sygnis S.L.U. Parque Científico de Madrid. Cantoblanco, Madrid, Spain.
authors.forename: Patricia
authors.initials: P
authors.lastname: Garrido
authors.affiliation: Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CIBERNED), ISCIII, Madrid, Spain.|Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Madrid, Spain.
authors.forename: Felix
authors.initials: F
authors.lastname: Hernandez
authors.affiliation: Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CIBERNED), ISCIII, Madrid, Spain.|Department of Cell Biology and Institute of Neurosciences, University of Barcelona, Barcelona, Spain.|Vall d'Hebrón Institut de Recerca (VHIR), Barcelona, Spain.|ICREA Academia, Barcelona, Spain.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Soriano
authors.affiliation: Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CIBERNED), ISCIII, Madrid, Spain.|Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Madrid, Spain.
authors.forename: Jesús
authors.initials: J
authors.lastname: Avila
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000483:Alleles; D000544:Alzheimer Disease / Q000097:blood / Q000235:genetics* / Q000378:metabolism*; D019295:Computational Biology / Q000379:methods*; D005260:Female; D006624:Hippocampus / Q000378:metabolism; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D020641:Polymorphism, Single Nucleotide; D017422:Sequence Analysis, DNA / Q000379:methods; D012984:Software; D054928:Validation Studies as Topic
keywords: Alzheimer’s disease; Sanger sequencing; Virmid software; identification of nucleotide variations; somatic mutations