Fluorescence-labeled methylation-sensitive amplified fragment length polymorphism (FL-MS-AFLP) analysis for quantitative determination of DNA methylation and demethylation status.
The PCR-based DNA fingerprinting method called the methylation-sensitive amplified fragment length polymorphism (MS-AFLP) analysis is used for genome-wide scanning of methylation status. In this study, we developed a method of fluorescence-labeled MS-AFLP (FL-MS-AFLP) analysis by applying a fluorescence-labeled primer and fluorescence-detecting electrophoresis apparatus to the existing method of MS-AFLP analysis. The FL-MS-AFLP analysis enables quantitative evaluation of more than 350 random CpG loci per run. It was shown to allow evaluation of the differences in methylation level of blood DNA of gastric cancer patients and evaluation of hypermethylation and hypomethylation in DNA from gastric cancer tissue in comparison with adjacent non-cancerous tissue.
Kageyama
Shinji
S
1st Department of Pathology, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi Ward, Hamamatsu, Shizuoka 431-3192, Japan.
Shinmura
Kazuya
K
Yamamoto
Hiroko
H
Goto
Masanori
M
Suzuki
Koichi
K
Tanioka
Fumihiko
F
Tsuneyoshi
Toshihiro
T
Sugimura
Haruhiko
H
D045744:Cell Line, Tumor; D018899:CpG Islands / Q000235:genetics; D016172:DNA Fingerprinting; D019175:DNA Methylation; D005456:Fluorescent Dyes; D006801:Humans; D007564:Japan; D008175:Lung Neoplasms / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques; D020411:Oligonucleotide Array Sequence Analysis; D012150:Polymorphism, Restriction Fragment Length; D013194:Staining and Labeling; D013274:Stomach Neoplasms / Q000235:genetics*
D005456:Fluorescent Dyes