AmpliconDesign - an interactive web server for the design of high-throughput targeted DNA methylation assays

Epigenetics. 2021 Sep;16(9):933-939. doi: 10.1080/15592294.2020.1834921. Epub 2020 Oct 24.

Abstract

Targeted analysis of DNA methylation patterns based on bisulfite-treated genomic DNA (BT-DNA) is considered as a gold-standard for epigenetic biomarker development. Existing software tools facilitate primer design, primer quality control or visualization of primer localization. However, high-throughput design of primers for BT-DNA amplification is hampered by limits in throughput and functionality of existing tools, requiring users to repeatedly perform specific tasks manually. Consequently, the design of PCR primers for BT-DNA remains a tedious and time-consuming process. To bridge this gap, we developed AmpliconDesign, a webserver providing a scalable and user-friendly platform for the design and analysis of targeted DNA methylation studies based on BT-DNA, e.g. deep amplicon bisulfite sequencing (ampBS-seq) or EpiTYPER MassArray. Core functionality of the web server includes high-throughput primer design and binding site validation based on in silico bisulfite-converted DNA sequences, prediction of fragmentation patterns for EpiTYPER MassArray, an interactive quality control as well as a streamlined analysis workflow for ampBS-seq.

Keywords: DNA methylation; analysis pipeline; bisulfite sequencing; epityper; massarray; primer design; targeted sequencing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Methylation*
  • Epigenomics
  • High-Throughput Nucleotide Sequencing
  • Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Software
  • Sulfites*

Substances

  • Sulfites

Grants and funding

This study has in part been supported by funds from Deutsche Krebshilfe [DKH project #70112574 to DBL] and from Deutsche Forschungsgemeinschaft [FOR2674, LI 2492/3-1 to DBL). PL was supported by AMPro Project of the Helmholtz Association [ZT00026].