High-Throughput, Amplicon-Based Sequencing of the CREBBP Gene as a Tool to Develop a Universal Platform-Independent Assay

PLoS One. 2015 Jun 9;10(6):e0129195. doi: 10.1371/journal.pone.0129195. eCollection 2015.

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.

MeSH terms

  • CREB-Binding Protein / genetics*
  • Computational Biology / methods
  • Exons / genetics
  • Genome, Human / genetics*
  • Genomics / methods
  • High-Throughput Screening Assays / methods*
  • Humans

Substances

  • CREB-Binding Protein
  • CREBBP protein, human

Associated data

  • SRA/SRP056569

Grants and funding

These authors have no support or funding to report.