Colorimetric detection of both total genomic and loci-specific DNA methylation from limited DNA inputs

Wee, Eugene J. H., Ngo, Thu Ha and Trau, Matt (2015) Colorimetric detection of both total genomic and loci-specific DNA methylation from limited DNA inputs. Clinical Epigenetics, 7 1: . doi:10.1186/s13148-015-0100-6

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Author Wee, Eugene J. H.
Ngo, Thu Ha
Trau, Matt
Title Colorimetric detection of both total genomic and loci-specific DNA methylation from limited DNA inputs
Journal name Clinical Epigenetics   Check publisher's open access policy
ISSN 1868-7075
1868-7083
Publication date 2015-07-11
Sub-type Article (original research)
DOI 10.1186/s13148-015-0100-6
Open Access Status File (Publisher version)
Volume 7
Issue 1
Total pages 9
Place of publication London, United Kingdom
Publisher BioMed Central
Collection year 2016
Language eng
Formatted abstract
Background: Aberrant DNA methylation marks are potential disease biomarkers, and detecting both total genomic
and gene-specific DNA methylation can aid in clinical decisions. While a plethora of methods exist in research,
simpler, more convenient alternatives are needed to enhance both routine diagnostics and research.

Results: Herein, we describe colorimetric assays using methyl-binding domain (MBD) proteins for rapid and
convenient evaluation of total genomic and gene-specific methylation from 50 ng or less DNA input in under 2 h.
As little as 5 % methylation differences can be detected and are enhanced by a novel MBD protocol for improved
specificity. Our assays could differentiate naïve from de-methylating drug-treated cells and detect the presence of a
methylated prostate cancer biomarker in the urine. Finally, the assay was evolved onto disposable screen-printed
electrodes for convenient detection of gene-specific methylation in urine.

Conclusions: Rapid MBD-based colorimetric and electrochemical approaches to detect DNA methylation from
limited samples were successfully demonstrated and applied to clinical samples. We envision that the ease, low
sample requirements and speed of these assays could have both clinical and research-wide applications.
Keyword MBD enrichment
DNA methylation
Colorimetric
Electrochemical
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

 
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Created: Wed, 15 Jul 2015, 09:28:47 EST by Eugene Wee on behalf of Aust Institute for Bioengineering & Nanotechnology