温度调制高效液相色谱法作为检测单核苷酸多态性工具的分析
[Analysis of temperature-modulated high-performance liquid chromatography as a tool for detection of single nucleotide polymorphism].
作者信息
Liao L, Meng H, Hou Y, Zhang S, Yan Y, Su Z, Li Y, Wu J, Zhang J
机构信息
Institute of Forensic Medicine, West China University of Medical Sciences, Chengdu, Sichuan, 610041 P. R. China.
出版信息
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2000 Jun;17(3):204-7.
OBJECTIVE
To establish and evaluate temperature-modulated high-performance liquid chromatography(TmHPLC) as a rapid and efficient technique of detecting single nucleotide polymorphism(SNP).
METHODS
Several TmHPLC parameters, such as the eluting gradient ranges and column temperatures used for analysing the amplicons have been studied. The size of DNA fragments suitable for analysis of SNP was studied. Also the sensitivity of the TmHPLC was evaluated.
RESULTS
The suitable chromatographic condition of TmHPLC was found out. The elution peaks of dNTP and primers could be separated from those of polymerase chain reaction products of the genomic DNA. The heterozygotes of SNP from the sequenced DNA fragment with 178bp, 269bp and 272bp could be distinguished from the homozygotes of SNP clearly.
CONCLUSION
The authors have established and evaluated the TmHPLC method, which is an efficient, sensitive and rapid method for screening out SNP in the DNA fragments of larger size.
目的
建立并评估温度调制高效液相色谱法（TmHPLC）作为一种快速有效的单核苷酸多态性（SNP）检测技术。
方法
研究了用于分析扩增子的几个TmHPLC参数，如洗脱梯度范围和柱温。研究了适合SNP分析的DNA片段大小。还评估了TmHPLC的灵敏度。
结果
找出了TmHPLC合适的色谱条件。dNTP和引物的洗脱峰可与基因组DNA聚合酶链反应产物的洗脱峰分离。来自178bp、269bp和272bp测序DNA片段的SNP杂合子可与SNP纯合子清楚区分。
结论
作者建立并评估了TmHPLC方法，这是一种高效、灵敏且快速的方法，用于筛选较大尺寸DNA片段中的SNP。
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