Ridanpää M, Burvall K, Zhang L H, Husgafvel-Pursiainen K, Onfelt A
Finnish Institute of Occupational Health, Helsinki.
Mutat Res. 1995 Jun;334(3):357-64. doi: 10.1016/0165-1161(95)90073-x.
Denaturing gradient gel electrophoresis (DGGE) and constant denaturant gel electrophoresis (CDGE) are methods based on sequence-determined melting characteristics of DNA and thus detect different types of single base changes in the amplified fragments. We have studied detection of 19 mutations in the human N-ras oncogene and 10 mutations in exon 3 of the Chinese hamster hypoxanthine-guanine phosphoribosyltransferase (hprt) gene using GC-clamped DGGE and CDGE. After allowing formation of heteroduplexes with the corresponding wild type sequence, all the mutations separated from the wild type in at least one concentration of the denaturants used in CDGE but two of the mutations in hprt exon 3 did not show separation in any of the DGGE runs. Melting behavior of the mutant fragments was dependent, as expected, on both the type and the location of a mutation. We describe conditions allowing separation of the mutations in the fewest possible DGGE and CDGE runs.
变性梯度凝胶电泳（DGGE）和恒定变性剂凝胶电泳（CDGE）是基于DNA序列决定的解链特性的方法，因此可以检测扩增片段中不同类型的单碱基变化。我们使用GC夹DGGE和CDGE研究了人类N-ras癌基因中的19个突变以及中国仓鼠次黄嘌呤-鸟嘌呤磷酸核糖转移酶（hprt）基因第3外显子中的10个突变的检测。在与相应的野生型序列形成异源双链体后，所有突变在CDGE中使用的至少一种变性剂浓度下与野生型分离，但hprt第3外显子中的两个突变在任何DGGE运行中均未显示分离。如预期的那样，突变片段的解链行为取决于突变的类型和位置。我们描述了在尽可能少的DGGE和CDGE运行中分离突变的条件。
