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PCR扩增配合以单链构象多态(SSCP)是检测特定DNA片段突变的较好方法。PCR扩增结果可能受到非特异扩增产物的影响,尤其是富含GC的序列。目前在反应中应用DM30,甲酰胺和核苷酸类似物7-deaza-2′-dGTP以提高特异性。本文作者发现在7-deaza-2′-dGTP与dGTP在3:1的情况下,可以大大地提高人IGF_2基因启动子区域某些片段的PCR扩增的特异性,然而,在接下去的PCR产物SSCP分析时却在凝胶中形成模糊的非特异区带。
PCR amplification combined with single-stranded conformational polymorphism (SSCP) is a better way to detect mutations in specific DNA fragments. PCR amplification results may be affected by non-specific amplification products, especially GC-rich sequences. DM30, formamide and the nucleotide analog 7-deaza-2’-dGTP are currently used in the reaction to enhance specificity. The authors found that in the case of 7-deaza-2’-dGTP and dGTP at 3: 1, the specificity of PCR amplification of certain fragments of the human IGF2 gene promoter region can be greatly enhanced, however, in the subsequent PCR The product was analyzed by SSCP in the gel to form a fuzzy non-specific zone.