高通量修饰的单链构象多态性分析及其在家族性高胆固醇血症突变检测中的应用。IFCC科学部:分子生物学技术委员会。

S E Humphries, V Gudnason, R Whittall, I N Day
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引用次数: 0

摘要

确定导致患者遗传疾病的特定突变是制定治疗和基于dna的筛查亲属试验的基本原理的框架。本文综述了单链构象多态性(SSCP)方法,该方法允许用特定引物和PCR扩增的DNA片段快速扫描任何序列变异。描述了该方法的一般原理,以及在制定最佳SSCP策略时必须考虑的主要因素,即PCR片段的长度和凝胶运行的温度。讨论了样品变性凝胶特性和DNA片段检测的选择。此外,还提出了一些用于高通量突变分析的修饰。应用这些技术筛选在低密度脂蛋白受体基因突变的患者家族性高胆固醇血症描述。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Single-strand conformation polymorphism analysis with high throughput modifications, and its use in mutation detection in familial hypercholesterolemia. The IFCC Scientific Division: Committee on Molecular Biology Techniques.

The identification of the specific mutation causing an inherited disease in a patient is the framework for the development of a rationale for therapy and of DNA-based tests for screening relatives. We present here a review of the single-strand conformational polymorphism (SSCP) method, which allows DNA fragments that have been amplified with specific primers and PCR to be scanned rapidly for any sequence variation. The general principles of the method are described, as are the major factors that must be considered in developing an optimal SSCP strategy, namely length of the PCR fragment and the temperature of the gel run. Options for sample denaturing gel characteristics and detection of DNA fragments are discussed. In addition, several modifications are presented that have been developed for high-throughput mutational analysis. The application of these techniques to screen for mutations in the LDL receptor gene in patients with familial hypercholesterolemia are described.

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