High-throughput, Amplicon-Based sequencing of the CREBBP gene as a tool to develop a universal platform-independent assay
| Autor: | Dietrich Herrmann, Monika Brüggemann, Michael Kneba, Sven Kuenzel, Henrik Knecht, Marc W. Fuellgrabe, Christiane Pott |
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| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Genetics
Cancer genome sequencing Multidisciplinary Massive parallel sequencing Genome Human Shotgun sequencing lcsh:R 2 base encoding lcsh:Medicine Computational Biology Hybrid genome assembly Exons Genomics Biology CREB-Binding Protein High-Throughput Screening Assays Single cell sequencing Humans lcsh:Q lcsh:Science Exome sequencing ABI Solid Sequencing Research Article |
| Zdroj: | PLoS One PLoS ONE PLoS ONE, Vol 10, Iss 6, p e0129195 (2015) |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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