Department of Pathology, University of Utah Health Sciences Center, Salt Lake City, UT 84132, USA.
Expert Rev Mol Diagn. 2010 Mar;10(2):219-40. doi: 10.1586/erm.09.84.
Among nucleic acid analytical methods, high-resolution melting analysis is gaining more and more attention. High-resolution melting provides simple, homogeneous solutions for variant scanning and genotyping, addressing the needs of today's overburdened laboratories with rapid turnaround times and minimal cost. The flexibility of the technique has allowed it to be adopted by a wide range of disciplines for a variety of applications. In this review we examine the broad use of high-resolution melting analysis, including gene scanning, genotyping (including small amplicon, unlabeled probe and snapback primers), sequence matching and methylation analysis. Four major application arenas are examined to demonstrate the methods and approaches commonly used in particular fields. The appropriate usage of high-resolution melting analysis is discussed in the context of known constraints, such as sample quality and quantity, with a particular focus placed on proper experimental design in order to produce successful results.
在核酸分析方法中,高分辨率熔解分析越来越受到关注。高分辨率熔解分析为变体扫描和基因分型提供了简单、均一的解决方案,满足了当今实验室负担过重的需求,具有快速周转时间和最小的成本。该技术的灵活性使其被广泛的学科应用于各种应用。在这篇综述中,我们考察了高分辨率熔解分析的广泛应用,包括基因扫描、基因分型(包括小扩增子、未标记探针和回扫引物)、序列匹配和甲基化分析。考察了四个主要的应用领域,以展示特定领域中常用的方法和方法。讨论了高分辨率熔解分析的适当使用,包括样品质量和数量等已知限制,特别强调了为获得成功结果而进行适当的实验设计。