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Mass Spectrometry Based Ultrasensitive DNA Methylation Profiling Using Target Fragmentation Assay
- Source :
- Analytical chemistry. 88(2)
- Publication Year :
- 2015
-
Abstract
- Efficient tools for profiling DNA methylation in specific genes are essential for epigenetics and clinical diagnostics. Current DNA methylation profiling techniques have been limited by inconvenient implementation, requirements of specific reagents, and inferior accuracy in quantifying methylation degree. We develop a novel mass spectrometry method, target fragmentation assay (TFA), which enable to profile methylation in specific sequences. This method combines selective capture of DNA target from restricted cleavage of genomic DNA using magnetic separation with MS detection of the nonenzymatic hydrolysates of target DNA. This method is shown to be highly sensitive with a detection limit as low as 0.056 amol, allowing direct profiling of methylation using genome DNA without preamplification. Moreover, this method offers a unique advantage in accurately determining DNA methylation level. The clinical applicability was demonstrated by DNA methylation analysis using prostate tissue samples, implying the potential of this method as a useful tool for DNA methylation profiling in early detection of related diseases.
- Subjects :
- 0301 basic medicine
Male
Nanotechnology
Computational biology
01 natural sciences
Genome
Mass Spectrometry
Analytical Chemistry
03 medical and health sciences
chemistry.chemical_compound
Humans
Epigenetics
Gene
Chemistry
010401 analytical chemistry
Prostate
Methylation
DNA
DNA Methylation
0104 chemical sciences
genomic DNA
030104 developmental biology
Magnetic Fields
DNA methylation
Illumina Methylation Assay
Subjects
Details
- ISSN :
- 15206882
- Volume :
- 88
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Analytical chemistry
- Accession number :
- edsair.doi.dedup.....e88a28058bad42d8da5efb769779bee5