DNA microarray: a high throughput approach for methylation detection.
We described a DNA microarray-based method combined with bisulphite treatment of DNA and regular PCR to examine hyper-methylation in promoter 1A of APC gene. A set of oligonucleotide probes were designed and immobilized on the aldehyde-coated glass slides for detecting the methylation pattern of 15 selected CpG sites in the region. The methylation status of 30 colorectal tumor samples have been examined by both of methylation-specific PCR (MS-PCR) and the present microarray method. The methylation pattern of the 15 CpG sites for the samples have been obtained with the microarray. A total of 19 samples out of 30 were methylated by microarray, in which five samples cannot be detected by MS-PCR due to the methylated CpG patterns not accordant to the MS-PCR primers. The detecting ratio for methylation of APC gene of colorectal tumor samples increased from 46.7% with MS-PCR to 63.3% with the microarray, which successfully demonstrated that DNA microarray-based method not only can obtained the methylation patterns for the related genes, but also decrease the false-negative results of methylation status by the conventional MS-PCR for the investigated genes.
Gao
L
L
Key Laboratory for Molecular and Biomolecular Electronics of Ministry of Education, Southeast University, Nanjing 210096, China.
Cheng
L
L
Zhou
J N
JN
Zhu
B L
BL
Lu
Z H
ZH
D025601:Adenomatous Polyposis Coli Protein / Q000235:genetics*; D001483:Base Sequence; D015179:Colorectal Neoplasms / Q000235:genetics; D019175:DNA Methylation; D015257:DNA-Cytosine Methylases; D006801:Humans; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic
D025601:Adenomatous Polyposis Coli Protein; C081573:DNA modification methylase SssI; D015257:DNA-Cytosine Methylases