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A next-generation sequencing method for overcoming the multiple gene copy problem in polyploid phylogenetics, applied to Poa grasses by Philippa C Griffin; Charles Robin; Ary A Hoffmann is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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## Background Polyploidy is important from a phylogenetic perspective because of its immense past impact on evolution and its potential future impact on diversification, survival and adaptation, especially in plants. Molecular population genetics studies of polyploid organisms have been difficult because of problems in sequencing multiple-copy nuclear genes using Sanger sequencing. This paper describes a method for sequencing a barcoded mixture of targeted gene regions using next-generation sequencing methods to overcome these problems. ## Results Using 64 3-bp barcodes, we successfully sequenced three chloroplast and two nuclear gene regions (each of which contained two gene copies with up to two alleles per individual) in a total of 60 individuals across 11 species of Australian __Poa__ grasses. This method had high replicability, a low sequencing error rate (after appropriate quality control) and a low rate of missing data. Eighty-eight percent of the 320 gene/individual combinations produced sequence reads, and >80% of individuals produced sufficient reads to detect all four possible nuclear alleles of the homeologous nuclear loci with 95% probability. We applied this method to

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Author

Philippa C Griffin; Charles Robin; Ary A Hoffmann

Publisher

BioMed Central; Springer (Biomed Central Ltd.); [London]: BioMed Central, c2003-; Springer Science and Business Media LLC; Society for Mining, Metallurgy and Exploration Inc.; Research Square (ISSN 1741-7007)

Published

2011

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)