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本文讨论了筛查未知突变的方法。1.固定在膜上的核酸与放射性探针杂交。Southern印迹法已广泛用于分子生物学,可以用于检出缺失、插入、重排这些大的基因变化。如由破坏的基因产生的mRNA能由Nothern印迹法检出,也许会对潜在的突变提供线索。2.以PCR为基础的突变检测,PCR在突变分析中最简单的应用是决定一段特异DNA的存在或缺失。多个DNA区域同时扩增,即所谓多重PCR,如在Duchenne肌营养不良中的应用。3.核酸异源双链的酶促切割,如果有了突变
This article discusses methods of screening for unknown mutations. 1. A nucleic acid immobilized on a membrane hybridizes to a radioactive probe. Southern blotting has been used extensively in molecular biology and can be used to detect large gene changes such as deletion, insertion, rearrangement. For example, mRNAs produced by disrupted genes can be detected by Nothern blotting, potentially providing clues to potential mutations. 2. PCR-based mutation detection, PCR The most simple application in mutation analysis is to determine the presence or absence of a specific DNA. Multiple DNA regions are simultaneously amplified, a so-called multiplex PCR, as in Duchenne muscular dystrophy. 3. Enzymatic cleavage of nucleic acid heteroduplexes, if mutated