DNA methylation analysis by MethyLight technology

被引:106
作者
Trinh, BN
Long, TI
Laird, PW
机构
[1] Univ So Calif, Keck Sch Med, Kenneth Norris Jr Comprehens Canc Ctr, Dept Surg, Los Angeles, CA 90089 USA
[2] Univ So Calif, Keck Sch Med, Kenneth Norris Jr Comprehens Canc Ctr, Dept Biochem & Mol Biol, Los Angeles, CA 90089 USA
关键词
CpG island; hypermethylation; DNA methylation; Taqman; real-time PCR; sodium bisulfite;
D O I
10.1006/meth.2001.1268
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
MethyLight is a sensitive, fluorescence-based real-time PCR technique that is capable of quantitating DNA methylation at a particular locus by using DNA oligonucleotides that anneal differentially to bisulfite-converted DNA according to the methylation status In the original genomic DNA. The use of three oligonucleotides (forward and reverse primers, and interpositioned probe) in MethyLight, any one or more of which can be used for methylation discrimination, allows for a high degree of specificity, sensitivity, and flexibility In methylation detection. (C) 2001 Elsevier Science (USA).
引用
收藏
页码:456 / 462
页数:7
相关论文
共 27 条
  • [1] Spreadsheet-based program for the analysis of DNA methylation
    Anbazhagan, R
    Herman, JG
    Enika, K
    Gabrielson, E
    [J]. BIOTECHNIQUES, 2001, 30 (01) : 110 - +
  • [2] DNA hypermethylation in tumorigenesis - epigenetics joins genetics
    Baylin, SB
    Herman, JG
    [J]. TRENDS IN GENETICS, 2000, 16 (04) : 168 - 174
  • [3] CPG-RICH ISLANDS AND THE FUNCTION OF DNA METHYLATION
    BIRD, AP
    [J]. NATURE, 1986, 321 (6067) : 209 - 213
  • [4] CLARK SJ, 1994, NUCLEIC ACIDS RES, V22, P2990, DOI 10.1093/nar/22.15.2990
  • [5] Eads CA, 2001, CANCER RES, V61, P3410
  • [6] Eads CA, 1999, CANCER RES, V59, P2302
  • [7] Eads CA, 2000, CANCER RES, V60, P5021
  • [8] MethyLight: a high-throughput assay to measure DNA methylation
    Eads, Cindy A.
    Danenberg, Kathleen D.
    Kawakami, Kazuyuki
    Saltz, Leonard B.
    Blake, Corey
    Shibata, Darryl
    Danenberg, Peter V.
    Laird, Peter W.
    [J]. NUCLEIC ACIDS RESEARCH, 2000, 28 (08) : 32
  • [9] A GENOMIC SEQUENCING PROTOCOL THAT YIELDS A POSITIVE DISPLAY OF 5-METHYLCYTOSINE RESIDUES IN INDIVIDUAL DNA STRANDS
    FROMMER, M
    MCDONALD, LE
    MILLAR, DS
    COLLIS, CM
    WATT, F
    GRIGG, GW
    MOLLOY, PL
    PAUL, CL
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (05) : 1827 - 1831
  • [10] Gonzalgo ML, 1997, CANCER RES, V57, P594