江西农业大学学报Issue(1)：169-175,7.
HRM分析方法中不同扩增片段、染料和仪器的比较
The Comparison of Amplified Fragments,Different Dyes and Instruments in the Analysis of HRM
摘要
Abstract
High resolution melting curve ( HRM ) technology is a new type of technology for genotyping and mutation scanning developing in recent years based on the technology of real⁃time fluorescence quantitative PCR.This study compared several causative agents of animal diseases,including Classical swine fever virus/Ei⁃meria species/Avain leukemia virus.The effects of different amplified regions of pathogens’ genomes(C202/ALV450/ En573 / C965),different fluorescent dyes(SYBR GreenⅠ/ LC Green / SYTO9)and different instruments(LightScanner96(Idaho) / LightCycler480(Roche) / Rotor⁃Gene Q(Qiagen)) on the HRM⁃based genotyping.Theexperimental data showed that HRM analysis was mainly affected by different amplified fragments and fluorescentdyes with different properties:nucleic acids with multiple melting range had higher specificity than that of singlenucleotide polymorphism(SNP)and were more suitable for differentiation analysis; difference in Tm on multiplemelting range influenced HRM results; saturated fluorescent dyes(LC Green and SYTO9) had higher resolutionthan unsaturated dye(SYBR GreenⅠ); concentration of fluorescent dye had a greater influence on the value ofTm; there was no significant difference in the HRM results from different PCR⁃HRM analyzers.关键词
高分辨率熔解曲线/单核苷酸多态性/荧光染料/熔解Tm/熔解曲线模拟Key words
HRM/SNP/fluorescent dye/melting Tm/imitating melting curve分类
农业科技引用本文复制引用
饶丹,郭鹏举,嘎利兵嘎,郭小权,张建峰,张春红,沈海燕,朱余军,刘志成,孙俊颖..HRM分析方法中不同扩增片段、染料和仪器的比较[J].江西农业大学学报,2015,(1):169-175,7.基金项目
国家自然科学基金资助项目（31260627）、广东省科技计划项目（2011B050700007，2012B050500013）、广东省重大专项（2012A020100001）、广东省动物防疫专项资金项目（粤农计［2013］31号）、江西省青年科学家培养对象项目（20122BCB23022）、广东省科技计划（星火计划）项目（2012A020602094）和国家国际科技合作专项 （）
