用温度调控高效液相色谱探索基因组单核苷酸多态性的方法研究  

Analysis of temperature modulated high-performance liquid chromatography as a tool for detection of single nucleotide polymorphism *

作　　者：廖林川[1] 孟海英[1] 侯一平[1] 张思仲 颜有仪[1] 苏智广 李英碧[1] 吴瑾[1] 张霁[1] 

机构地区：[1]华西医科大学法医学院附属第一医院,成都610041 [2]医学遗传研究室

出　　处：《中华医学遗传学杂志》2000年第3期204-207,共4页Chinese Journal of Medical Genetics

基　　金：国家自然科学基金!(3999342 0 )

摘　　要：目的　优化并评估一种新的探索基因组单核苷酸多态 (single nucleotide polymorphism,SNP)的温度调控高效液相色谱技术。方法　从洗脱方式、洗脱温度、检测器流通池规格、适应片段大小、检测灵敏度等方面 ,探讨适合探索基因组 SNP的温度调控高效液相色谱技术参数及实用价值。结果　找到在普通高效液相色谱仪上检测 SNP的具有普遍意义的色谱条件。 d NTP、PCR引物峰与被检测 PCR产物峰完全分离 ,纯合子和杂合子样本能有效区分。建立起在普通高效液相色谱仪上进行快速筛选突变型DNA的温度调控高效液相色谱法。结论　温度调控高效液相色谱法是一种高效、灵敏、操作简便、对较大片段具有普遍适用意义的探索基因组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 178 bp, 269 bp and 272 bp 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.

关 键 词：温度调控 高效液相色谱 单核苷酸多态性 基因组 

分 类 号：Q75[生物学—分子生物学]