| 苹果不同HRM反应体系分析效果评价 |
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| 引用本文： | 白牡丹,王彩虹,殷豪,田义轲,李节法.苹果不同HRM反应体系分析效果评价[J].分子植物育种,2012(1):115-120. |
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| 作者姓名： | 白牡丹 王彩虹 殷豪 田义轲 李节法 |
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| 作者单位： | 青岛农业大学园林园艺学院 |
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| 基金项目： | 山东省良种工程项目和青岛市科技计划基础研究项目(11-2-4-5-(4)-jch)共同资助 |
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| 摘 要： | 高分辨率熔解曲线(high resolution melting,HRM)分析是一种新型的DNA多态性快速筛查技术。本研究以苹果品种"富士"和"舞姿"为试材,用来自苹果基因组的SSR标记CH03d11对不同反应体积、不同DNA浓度以及不同PCR退火程序下的HRM分型效果进行了测试与评估。结果表明:采用5μL反应体积可以获得与10μL或20μL反应体积相同的检测效果。在5μL反应体积中,DNA浓度小到0.25ng/μL时,PCR扩增及随后的HRM分析效果依然良好。另外,研究表明,采用降落PCR扩增模式也能取得良好的HRM多态性检测结果。
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| 关 键 词： | 苹果(Malus domestica) HRM 反应体积 DNA模板浓度 降落PCR |
Evaluation of Different Reaction Systems for HRM Analysis in Apple |
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| i Mudan,g Caihong,n Hao,n Yike,i Jiefa.Evaluation of Different Reaction Systems for HRM Analysis in Apple[J].Molecular Plant Breeding,2012(1):115-120. |
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| Authors: | i Mudan g Caihong n Hao n Yike i Jiefa |
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| Institution: | College of Landscape and Horticulture,Qingdao Agricultural University,Qindao,266109 |
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| Abstract: | High resolution melting(HRM) analysis is a newly developed method for fast DNA polymorphism detection.HRM analysis under different reaction volumes,DNA concentrations or polymerase chain reaction(PCR) annealing programs were evaluated for genotyping in apple with cultivar ’Fuji’ and ’Telamon’.The result indicated that 5 μL reaction volume was as efficient as 10 μL or 20 μL for revealed the polymorphism of SSR(simple sequence repeat) marker CH03d11,which derived from apple genome,between the two cultivars.Therefore,even DNA concentration as small as 0.25 ng/μL was good enough for PCR amplification and the following HRM detection under a 5 μL reaction volume.Additional study demonstrated that a touchdown PCR program could also performed very well in HRM analysis for polymorphism detection. |
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| Keywords: | Apple(Malus domestica) HRM Reaction volume DNA template concentration Touchdown PCR |
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