| 一种改良的亚硫酸氢盐修饰微量DNA的甲基化分析方法 |
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| 引用本文： | 冯苏妹,杨磊,吕嘉春,纪卫东. 一种改良的亚硫酸氢盐修饰微量DNA的甲基化分析方法[J]. 现代保健, 2008, 0(32): 9-11 |
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| 作者姓名： | 冯苏妹 杨磊 吕嘉春 纪卫东 |
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| 作者单位： | 广州医学院化学致癌研究所,510182 |
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| 基金项目： | 国家自然科学基金项目（No.30671746）,广东省自然科学基金项目（No.07003055） |
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| 摘 要： | 目的建立一种改良的亚硫酸氢盐修饰微量DNA的甲基化分析方法。方法应用低熔点琼脂糖包埋不同含量的DNA，进行亚硫酸氢盐修饰，通过MGMT基因甲基化与非甲基化特异PCR的扩增，与传统的亚硫酸氢盐修饰法进行分析比较。结果改良法对于低至15．625ng的DNA修饰均可获得MGMT基因甲基化扩增产物，而传统法对于62．5ng及其以下的DNA修饰难以获得MGMT基因甲基化扩增产物。结论改良的亚硫酸氢盐修饰方法可有效提高甲基化特异PCR的灵敏度，该方法为微量DNA的甲基化分析提供了一种较理想的研究手段。
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| 关 键 词： | 亚硫酸氢盐修饰 微量DNA 甲基化特异PCR(MSP) |
A modified method of bisulfite modification of trace DNA methylation analysis. |
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| Affiliation: | FENG Su - mei, YANG Lei, LU Jia - chun, JI Wei - dong. (Institute of Chemical Carcinogenesis, Guangzhou Medical College. Guangzhou 510182, China.) |
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| Abstract: | Objective Develop a modified method of bisulfite modification of trace DNA for analysis of DNA methylation. Method Series contents DNA embedded in low melting point agar were modified with bisulfite, amplified by methylation specific PCR for MGMT gene, which compared with the traditional method of bisulfite modification. Results MSP for MGMT gene were amplified successfully in 15. 526ng DNA modifled with bisuffite by the modified method, however amplifications of MSP failed in or under 62.5ng DNA modified by the traditional method. Conclusion The modified method of bisulfite modification could improve sensitivity of MSP effectively, and it provides a better tool for analyse trace DNA methylation. |
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| Keywords: | Bisufite modification Trace DNA Methylation spectific PCR（MSP） |
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