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  High-throughput, Amplicon-Based sequencing of the CREBBP gene as a tool to develop a universal platform-independent assay

Fuellgrabe, M. W., Herrmann, D., Knecht, H., Kuenzel, S., Kneba, M., Pott, C., et al. (2015). High-throughput, Amplicon-Based sequencing of the CREBBP gene as a tool to develop a universal platform-independent assay. PLoS One, 0129195. doi:10.1371/journal.pone.0129195.

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Fuellgrabe, Marc W., Autor
Herrmann, Dietrich, Autor
Knecht, Henrik, Autor
Kuenzel, Sven1, Autor           
Kneba, Michael, Autor
Pott, Christiane, Autor
Brüggemann, Monika, Autor
Affiliations:
1Department Evolutionary Genetics, Max Planck Institute for Evolutionary Biology, Max Planck Society, ou_1445635              

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 Zusammenfassung: High-throughput sequencing technologies are widely used to analyse genomic variants or rare mutational events in different fields of genomic research, with a fast development of new or adapted platforms and technologies, enabling amplicon-based analysis of single target genes or even whole genome sequencing within a short period of time. Each sequencing platform is characterized by well-defined types of errors, resulting from different steps in the sequencing workflow. Here we describe a universal method to prepare amplicon libraries that can be used for sequencing on different high-throughput sequencing platforms. We have sequenced distinct exons of the CREB binding protein (CREBBP) gene and analysed the output resulting from three major deep-sequencing platforms. platform-specific errors were adjusted according to the result of sequence analysis from the remaining platforms. Additionally, bioinformatic methods are described to determine platform dependent errors. Summarizing the results we present a platform-independent cost-efficient and timesaving method that can be used as an alternative to commercially available sample-preparation kits.

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Sprache(n): eng - English
 Datum: 2015-03-092015-04-072015-06-09
 Publikationsstatus: Online veröffentlicht
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 Ort, Verlag, Ausgabe: -
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 Art der Begutachtung: Expertenbegutachtung
 Identifikatoren: DOI: 10.1371/journal.pone.0129195
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Titel: PLoS One
Genre der Quelle: Zeitschrift
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Ort, Verlag, Ausgabe: San Francisco, CA : Public Library of Science
Seiten: 12 S. Band / Heft: - Artikelnummer: 0129195 Start- / Endseite: - Identifikator: ISSN: 1932-6203
CoNE: https://pure.mpg.de/cone/journals/resource/1000000000277850