Division of Genetics, Department of Biology, Faculty of Science, The University of Isfahan, Hezarjerib St., Isfahan, IR, Iran.
Appl Biochem Biotechnol. 2010 Mar;160(3):927-31. doi: 10.1007/s12010-009-8595-y. Epub 2009 Mar 31.
The single-strand conformation polymorphism (SSCP), accompanied by sequencing, is a useful methods for identifying mutations in a DNA fragment. In this study, we have developed a modified SSCP with the aid of sodium bisulfite treatment. The corresponding PCR products for exon 3 of Hb gene were sequenced and samples with homozygote and heterozygote single nucleotide substitutions were identified. The PCR products were treated with sodium bisulfite, which deaminates all the cytosine residues. The reaction mixture was then analyzed on non-denaturing polyacrylamide gels. The modified method, which is called deaminated SSCP (DSSCP), was applied successfully in analysis of mutations in the beta-globin gene at positions relevant to codon 6. DSSCP is a very effective and reproducible method providing clear results that are easy to interpret without the involvement of radioactivity.
单链构象多态性（SSCP）结合测序是一种用于鉴定 DNA 片段中突变的有用方法。在这项研究中，我们借助亚硫酸氢钠处理开发了一种改良的 SSCP。对 Hb 基因外显子 3 的相应 PCR 产物进行测序，并鉴定出纯合子和杂合子单核苷酸替换的样本。用亚硫酸氢钠处理 PCR 产物，使所有胞嘧啶残基脱氨基。然后将反应混合物在非变性聚丙烯酰胺凝胶上进行分析。这种改良的方法称为脱氨基 SSCP（DSSCP），成功地应用于分析与密码子 6 相关位置的β-球蛋白基因的突变。DSSCP 是一种非常有效且可重复的方法，提供清晰的结果，易于解释，无需涉及放射性。
