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Targeted DNA Methylation Analysis by Next-generation Sequencing
Published on: February 24, 2015
Rapid and sensitive detection of CpG-methylation using methyl-binding (MB)-PCR
Claudia Gebhard1, Lucia Schwarzfischer, Thu Hang Pham
1Department of Hematology and Oncology, University Hospital, 93042, Regensburg, Germany.
Nucleic Acids Research
|July 11, 2006
Summary
We developed methyl-binding PCR (MB-PCR), a novel method to detect methylated DNA. This technique identified aberrant hypermethylation of the ICSBP gene in acute myeloid leukemia.
Area of Science:
- Molecular Biology
- Cancer Genetics
- Epigenetics
Background:
- CpG island methylation silences tumor suppressor genes in cancer.
- Detecting DNA methylation is crucial for cancer research.
Purpose of the Study:
- To develop a novel, sensitive technique for detecting CpG-methylated DNA.
- To identify novel methylated genes in acute myeloid leukemia.
Main Methods:
- Developed methyl-binding (MB)-PCR, a technique using a recombinant protein to capture methylated DNA.
- MB-PCR selectively isolates methylated DNA fragments for detection via gene-specific PCR.
- This method avoids bisulfite treatment and methylation-sensitive restriction.
Main Results:
- MB-PCR is a quick, simple, and highly sensitive method for detecting methylated DNA, especially in limited tumor samples.
- Applied MB-PCR to analyze methylation status of tumor suppressor genes.
- Identified aberrant hypermethylation of the ICSBP gene in acute myeloid leukemia.
Conclusions:
- MB-PCR is an effective tool for detecting DNA methylation.
- Aberrant hypermethylation of the ICSBP gene is implicated in acute myeloid leukemia development.
