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

Fuellgrabe, Marc W.; Herrmann, Dietrich; Knecht, Henrik; Kuenzel, Sven; Kneba, Michael; Pott, Christiane; Brüggemann, Monika

doi:10.1371/journal.pone.0129195

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

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Kuenzel, Sven
Department Evolutionary Genetics, Max Planck Institute for Evolutionary Biology, Max Planck Society;

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Citation

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.

Cite as: https://hdl.handle.net/11858/00-001M-0000-0027-BA30-B

Abstract

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.