高阶导数技术在重新结合的重复DNA高分辨率热变性图谱分析中的应用。
Application of higher derivative techniques to analysis of high-resolution thermal denaturation profiles of reassociated repetitive DNA.
作者信息
Cuellar R E, Ford G A, Briggs W R, Thompson W F
出版信息
Proc Natl Acad Sci U S A. 1978 Dec;75(12):6026-30. doi: 10.1073/pnas.75.12.6026.
Abstract
We have analyzed high-resolution denaturation profiles of reassociated repetitive DNA sequences by using a combination of higher derivative analysis and curve-fitting techniques. Procedures originally used for resolution of components in complex absorption spectra were found to be applicable to high-resolution analysis of melting profiles of reassociated repetitive DNA sequences from pea DNA. Under conditions that eliminate the base composition effect on thermal stability (2.4 M tetraethylammonium chloride), such an anlysis can distinquish "thermal classes" of repetitive DNA duplexes exhibiting different amounts of base pair mismatch. Only a single thermal class is observed in reassociated Escherichia coli DNA whereas at least five classes can be reproducibly distinguished in pea and mung bean DNAs.
摘要
我们通过结合高阶导数分析和曲线拟合技术，分析了重退火重复DNA序列的高分辨率变性图谱。最初用于解析复杂吸收光谱中各成分的方法，被发现适用于豌豆DNA重退火重复DNA序列解链图谱的高分辨率分析。在消除碱基组成对热稳定性影响的条件下（2.4 M四乙基氯化铵），这样的分析能够区分表现出不同碱基对错配量的重复DNA双链体的“热类别”。在重退火的大肠杆菌DNA中仅观察到一个热类别，而在豌豆和绿豆DNA中至少可以重复区分出五个类别。
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