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。
Objective Development of a modified method of bisulfite modification of micro - quantity of 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 modified with bisulfite 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 effectively sensitivity of MSP, and it provides a better tool for micro-quantity of DNA for analysis of DNA methylation.
基金
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）
