THE SENSITIVITY OF SINGLE-STRAND CONFORMATION POLYMORPHISM ANALYSIS FOR THE DETECTION OF SINGLE BASE SUBSTITUTIONS
THE SENSITIVITY OF SINGLE-STRAND CONFORMATION POLYMORPHISM ANALYSIS FOR THE DETECTION OF SINGLE BASE SUBSTITUTIONS
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DOI：
10.1006/geno.1993.1193
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发表时间：
1993-05-01
期刊：
影响因子：
4.4
通讯作者：
STONE, EM
中科院分区：
文献类型：
--
作者：
SHEFFIELD, VC;BECK, JS;STONE, EM
单链构象多态性（SSCP）分析已被证明是检测单碱基取代的一种简单有效的技术。我们使用SSCP分析了不同大小PCR产物中包含的29个小鼠珠蛋白突变，27个p53突变和8个视紫红质突变。我们的研究结果表明，突变类型（转变与翻转）在SSCP分析中是否检测到突变并没有起主要作用。在确定是否检测到突变时，碱基取代的位置比精确的碱基取代更重要。我们报告说，SSCP的敏感性随着DNA片段的大小而急剧变化。SSCP检测敏感碱基取代的最佳片段大小约为150 bp。我们的结果表明，在进行SSCP检测突变时，需要保持PCR片段的小尺寸。当检测每个序列变异的需求不那么重要时，可以在筛选多态性时分析较大的片段。
Single-strand conformation polymorphism (SSCP) analysis has proven to be a simple and effective technique for the detection of single base substitutions. We have used SSCP to analyze 29 mouse globin mutations, 27 p53 mutations, and 8 rhodopsin mutations contained within different size PCR products. Our results indicate that the type of mutation (transition versus transversion) did not play a major role in determining whether a mutation was detected by SSCP analysis. The position of the base substitution was more important than the precise base substitution in determining whether a mutation was detected. We report that SSCP sensitivity varies dramatically with the size of the DNA fragment being analyzed. The optimal size fragment for sensitive base substitution detection by SSCP is approximately 150 bp. Our results illustrate the need to keep the size of the PCR fragment small when performing SSCP to detect mutations. Larger fragments can be analyzed when screening for polymorphisms when the need to detect every sequence variation is not as critical.