| Literature DB >> 25129690 |
Paula J P de Vree1, Elzo de Wit2, Mehmet Yilmaz3, Monique van de Heijning3, Petra Klous3, Marjon J A M Verstegen4, Yi Wan4, Hans Teunissen4, Peter H L Krijger4, Geert Geeven4, Paul P Eijk5, Daoud Sie5, Bauke Ylstra5, Lorette O M Hulsman6, Marieke F van Dooren6, Laura J C M van Zutven6, Ans van den Ouweland6, Sjef Verbeek7, Ko Willems van Dijk7, Marion Cornelissen8, Atze T Das8, Ben Berkhout8, Birgit Sikkema-Raddatz9, Eva van den Berg9, Pieter van der Vlies9, Desiree Weening9, Johan T den Dunnen10, Magdalena Matusiak11, Mohamed Lamkanfi11, Marjolijn J L Ligtenberg12, Petra ter Brugge13, Jos Jonkers13, John A Foekens14, John W Martens14, Rob van der Luijt15, Hans Kristian Ploos van Amstel15, Max van Min3, Erik Splinter3, Wouter de Laat16.
Abstract
Despite developments in targeted gene sequencing and whole-genome analysis techniques, the robust detection of all genetic variation, including structural variants, in and around genes of interest and in an allele-specific manner remains a challenge. Here we present targeted locus amplification (TLA), a strategy to selectively amplify and sequence entire genes on the basis of the crosslinking of physically proximal sequences. We show that, unlike other targeted re-sequencing methods, TLA works without detailed prior locus information, as one or a few primer pairs are sufficient for sequencing tens to hundreds of kilobases of surrounding DNA. This enables robust detection of single nucleotide variants, structural variants and gene fusions in clinically relevant genes, including BRCA1 and BRCA2, and enables haplotyping. We show that TLA can also be used to uncover insertion sites and sequences of integrated transgenes and viruses. TLA therefore promises to be a useful method in genetic research and diagnostics when comprehensive or allele-specific genetic information is needed.Entities:
Mesh：
Year: 2014 PMID： 25129690 DOI： 10.1038/nbt.2959
Source DB: PubMed Journal: Nat Biotechnol ISSN： 1087-0156 Impact factor: 54.908