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Updated: Feb 10, 2026
13:47
Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution
Published on: February 24, 2015
26.5K
A Rapid and Robust Protocol for Reduced Representation Bisulfite Sequencing in Multiple Myeloma
Samrat Roy Choudhury1, Brian A Walker2
1Department of Internal Medicine, Myeloma Institute, University of Arkansas for Medical Sciences, Little Rock, AR, USA.
Methods in Molecular Biology (Clifton, N.J.)
|May 26, 2018
Summary
We optimized reduced representation bisulfite sequencing (RRBS) for low DNA input, enabling efficient whole genome DNA methylation mapping in clinical samples within three days.
Area of Science:
- Epigenetics
- Genomics
- Molecular Biology
Background:
- Reduced representation bisulfite sequencing (RRBS) is a cost-effective method for whole genome DNA methylation analysis.
- Clinical samples, particularly myeloma, often have limited DNA input, posing challenges for traditional RRBS protocols.
Purpose of the Study:
- To develop a modified RRBS protocol optimized for low DNA input (as low as 100 ng).
- To streamline the RRBS workflow for faster processing of clinical samples.
Main Methods:
- Combined end-repair and A-tailing steps.
- Introduced a bead-based method for rapid size selection of library molecules.
- Optimized the protocol for myeloma samples with low DNA concentrations.
Main Results:
- The modified RRBS protocol significantly reduces the input DNA requirement.
- The entire protocol, including overnight digestion, can be completed in three days.
- The method is effective for DNA methylation profiling in samples with very low DNA concentrations.
Conclusions:
- The enhanced RRBS protocol provides a comprehensive and economical approach for DNA methylation analysis.
- This optimized method is broadly applicable to various clinical samples with limited DNA availability.
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