DNA测序与熔解曲线。
DNA sequencing and melting curve.
作者信息
Azbel M Y
出版信息
Proc Natl Acad Sci U S A. 1979 Jan;76(1):101-5. doi: 10.1073/pnas.76.1.101.
The dependence of DNA absorbance (for light at about 260 nm) on temperature is related to a specific DNA sequence structure in the vicinity of DNA thermal denaturation (the so-called DNA melting or coiling). A straightforward analysis of the experimental DNA melting curve allows us to determine the lengths, the A+T content, and the location in DNA of certain domains. In the case of a specific DNA fragmentation, the order of fragments in DNA can be learned from this analysis, nondestructively and quickly, without fractionating the fragments and other methods of fragmentation. If the DNA nucleotide sequence is known except for some sites and uncertain portions, the analysis determines these sites and the accuracy of the sequence at the portions. This information may complement exact methods of DNA sequencing. The proposed analysis is applied to bacteriophage phiX174, whose melting curve is known. The results are compared to and found to be in an excellent agreement with the known phiX174 nucleotide sequence.
DNA吸光度（对于约260nm波长的光）对温度的依赖性与DNA热变性（所谓的DNA解链或卷曲）附近的特定DNA序列结构有关。对实验性DNA解链曲线进行直接分析，能够确定某些结构域的长度、A+T含量以及在DNA中的位置。对于特定的DNA片段化情况，无需对片段进行分级分离和其他片段化方法，通过该分析就能无损且快速地得知DNA中片段的顺序。如果除了某些位点和不确定部分外，DNA核苷酸序列已知，那么该分析可以确定这些位点以及这些部分序列的准确性。此信息可补充DNA测序的精确方法。将所提出的分析方法应用于已知解链曲线的噬菌体phiX174。结果与已知的phiX174核苷酸序列进行比较，发现两者吻合度极高。
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