Sadri R, Hornsby P J
Huffington Center on Aging and Department of Cell Biology, Baylor College of Medicine, Houston, TX 77030, USA.
Nucleic Acids Res. 1996 Dec 15;24(24):5058-9. doi: 10.1093/nar/24.24.5058.
Bisulfite converts non-methylated cytosine in DNA to uracil leaving 5-methylcytosine unaltered. Here, predicted changes in restriction enzyme sites following reaction of genomic DNA with bisulfite and amplification of the product by the polymerase chain reaction (PCR) were used to assess the methylation of CpG sites. This procedure differs from conventional DNA methylation analysis by methylation-sensitive restriction enzymes because it does not rely on an absence of cleavage to detect methylated sites, the two strands of DNA produce different restriction enzyme sites and may be differentially analyzed, and closely related sequences may be separately analyzed by using specific PCR primers.
亚硫酸氢盐可将DNA中的非甲基化胞嘧啶转化为尿嘧啶，而5-甲基胞嘧啶则保持不变。在此，通过基因组DNA与亚硫酸氢盐反应后预测的限制性酶切位点变化以及聚合酶链反应（PCR）对产物的扩增来评估CpG位点的甲基化情况。该方法不同于传统的利用甲基化敏感限制性酶进行的DNA甲基化分析，因为它不依赖于未切割来检测甲基化位点，DNA的两条链会产生不同的限制性酶切位点且可进行差异分析，并且通过使用特异性PCR引物可对密切相关的序列进行单独分析。
