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Related Experiment Video

Updated: May 16, 2026

Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution
13:47

Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution

Published on: February 24, 2015

Technical considerations for reduced representation bisulfite sequencing with multiplexed libraries.

Aniruddha Chatterjee1, Euan J Rodger, Peter A Stockwell

  • 1Department of Pathology, Dunedin School of Medicine, University of Otago, 270 Great King Street, Dunedin 9054, New Zealand.

Journal of Biomedicine & Biotechnology
|November 30, 2012
PubMed
Summary
This summary is machine-generated.

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This study optimizes multiplexed reduced representation bisulfite sequencing (RRBS) for Illumina platforms. Modifications enable efficient DNA methylation analysis at single-nucleotide resolution using high-throughput sequencing.

Area of Science:

  • Genomics
  • Epigenetics
  • Next-Generation Sequencing

Background:

  • Reduced representation bisulfite sequencing (RRBS) is crucial for studying DNA methylation at single-nucleotide resolution.
  • Illumina sequencing technology offers high throughput but presents challenges for multiplexed RRBS libraries.
  • Optimizing multiplexed library preparation is essential for efficient genomic analysis.

Purpose of the Study:

  • To describe modifications for sequencing multiplexed RRBS libraries on the Illumina HiSeq 2000.
  • To present a bioinformatics pipeline for analyzing multiplexed RRBS data.
  • To provide strategies for troubleshooting and optimizing multiplexed RRBS sequencing.

Main Methods:

  • Modified the RRBS library preparation protocol for multiplexed samples.

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Targeted DNA Methylation Analysis by Next-generation Sequencing

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Last Updated: May 16, 2026

Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution
13:47

Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution

Published on: February 24, 2015

Enrichment of mRNA and Bisulfite-mRNA Library Preparation for Next-Generation Sequencing
06:57

Enrichment of mRNA and Bisulfite-mRNA Library Preparation for Next-Generation Sequencing

Published on: July 7, 2023

Targeted DNA Methylation Analysis by Next-generation Sequencing
08:38

Targeted DNA Methylation Analysis by Next-generation Sequencing

Published on: February 24, 2015

  • Utilized Illumina HiSeq 2000 for high-throughput sequencing.
  • Developed a bioinformatics pipeline for processing and quality assessment of bisulfite-converted reads.

    • Main Results:

    • Successfully sequenced multiplexed RRBS libraries on a single flow cell lane.
    • Achieved an average of 42 million paired-end reads per sample.
    • Demonstrated high unique mapping efficiency to the human reference genome.

    Conclusions:

    • The described modifications and bioinformatics pipeline optimize multiplexed RRBS sequencing on Illumina platforms.
    • This approach enhances the efficiency and throughput of DNA methylation studies.
    • Provides a valuable roadmap for researchers utilizing multiplexed RRBS libraries.