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BAL31-NGS approach for identification of telomeres de novo in large genomes

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PEŠKA Vratislav ŠEDĚNKA Zdeňka FAJKUS Petr FAJKUS Jiří

Rok publikování 2017
Druh Článek v odborném periodiku
Časopis / Zdroj Methods
Fakulta / Pracoviště MU

Středoevropský technologický institut

Citace
www https://www.sciencedirect.com/science/article/pii/S1046202316302900?via%3Dihub
Doi http://dx.doi.org/10.1016/j.ymeth.2016.08.017
Klíčová slova Telomere; NGS; BAL31; RepeatExplorer; Tandem Repeats Finder; Tandem Repeats Merger
Popis This article describes a novel method to identify as yet undiscovered telomere sequences, which combines next generation sequencing (NGS) with BAL31 digestion of high molecular weight DNA. The method was applied to two groups of plants: i) dicots, genus Cestrum, and ii) monocots, Allium species (e.g. A. ursinum and A. cepa). Both groups consist of species with large genomes (tens of Gb) and a low number of chromosomes (2n =14-16), full of repeat elements. Both genera lack typical telomeric repeats and multiple studies have attempted to characterize alternative telomeric sequences. However, despite interesting hypotheses and suggestions of alternative candidate telomeres (retrotransposons, rDNA, satellite repeats) these studies have not resolved the question. In a novel approach based on the two most general features of eukaryotic telomeres, their repetitive character and sensitivity to BAL31 nuclease digestion, we have taken advantage of the capacity and current affordability of NGS in combination with the robustness of classical BAL31 nuclease digestion of chromosomal termini. While representative samples of most repeat elements were ensured by low-coverage (less than 5%) genomic shot-gun NGS, candidate telomeres were identified as under-represented sequences in BAL31-treated samples. (C) 2016 Elsevier Inc. All rights reserved.
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