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    Aiming off the target: recycling target capture sequencing reads for investigating repetitive DNA

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    Authors
    Costa, Lucas
    Marques, André
    Buddenhagen, Chris
    Thomas, William Wayt
    Huettel, Bruno
    Schubert, Veit
    Dodsworth, Steven
    Houben, Andreas
    Souza, Gustavo
    Pedrosa-Harand, Andrea
    Issue Date
    2021-05-29
    Subjects
    high-throughput sequencing
    repetitive DNA
    
    Metadata
    Show full item record
    Abstract
    With the advance of high-throughput sequencing (HTS), reduced-representation methods such as target capture sequencing (TCS) emerged as cost-efficient ways of gathering genomic information, particularly from coding regions. As the off-target reads from such sequencing are expected to be similar to genome skimming (GS), we assessed the quality of repeat characterization in plant genomes using this data. Repeat composition obtained from TCS datasets of five Rhynchospora (Cyperaceae) species were compared with GS data from the same taxa. In addition, a FISH probe was designed based on the most abundant satellite found in the TCS dataset of Rhynchospora cephalotes. Finally, repeat-based phylogenies of the five Rhynchospora species were constructed based on the GS and TCS dataset and the topologies were compared with a gene-alignment based phylogenetic tree. All the major repetitive DNA families were identified in TCS, including repeats that showed abundances as low as 0.01% in the GS data. Rank correlation between GS and TCS repeat abundances were moderately high (r = 0.58-0.85), increasing after filtering out the targeted loci from the raw TCS reads (r = 0.66-0.92). Repeat data obtained by TCS was also reliable to develop a cytogenetic probe of a new variant of the holocentromeric satellite Tyba. Repeat-based phylogenies from TCS data were congruent with those obtained from GS data and the gene-alignment tree. Our results show that off-target TCS reads can be recycled to identify repeats for cyto- and phylogenomic investigations. Given the growing availability of TCS reads, driven by global phylogenomic projects, our strategy represents a way to recycle genomic data and contribute to a better characterization of plant biodiversity.
    Citation
    Costa L, Marques A, Buddenhagen C, Thomas WW, Huettel B, Schubert V, Dodsworth S, Houben A, Souza G, Pedrosa-Harand A (2021) 'Aiming off the target: recycling target capture sequencing reads for investigating repetitive DNA', Annals of Botany, 128 (7), pp.835-848.
    Publisher
    Oxford University Press
    Journal
    Annals of Botany
    URI
    http://hdl.handle.net/10547/625011
    DOI
    10.1093/aob/mcab063
    PubMed ID
    34050647
    PubMed Central ID
    PMC8577205
    Additional Links
    https://academic.oup.com/aob/advance-article-abstract/doi/10.1093/aob/mcab063/6288108
    Type
    Article
    Language
    en
    ISSN
    0305-7364
    EISSN
    1095-8290
    ae974a485f413a2113503eed53cd6c53
    10.1093/aob/mcab063
    Scopus Count
    Collections
    Biomedical and biological science

    entitlement

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