Digital High-Resolution Melt Platform for Rapid and Parallelized Molecule-by-Molecule Genetic Profiling.
Digital High-Resolution Melt Platform for Rapid and Parallelized Molecule-by-Molecule Genetic Profiling.
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用于快速并行逐分子遗传分析的数字高分辨率熔解平台。
DOI：
10.1109/embc.2018.8513609
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Wang,Tza-HueilJeffr
中科院分区：
文献类型：
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作者：
OrKeefe,ChristineM;Wang,Tza-HueilJeffr
这项工作提出了一种微流体数字高分辨率熔解平台，用于在逐个分子的基础上对位点特异性序列变异进行绝对定量和灵敏检测。该平台通过数字化 4096 1-nL 阵列微流体装置中的模板分子并观察温度升高过程中序列依赖性荧光变化，提供了一种评估数百至数千个单个 DNA 拷贝的简便方法。该平台的分析能力在多种应用中得到了证明，例如数字化验表征、DNA 甲基化异质性的检测和评估，以及以低至背景分子目标 0.0005% 的频率检测稀有生物标志物。
This work presents a microfluidic Digital High-Resolution Melt platform for absolute quantitation and sensitive detection of locus-specific sequence variations on a molecule-by-molecule basis. The platform provides a facile means for assessment of hundreds to thousands of single DNA copies by digitizing template molecules in a 4096 1-nL array microfluidic device and observing the sequence-dependent fluorescence changes during temperature ramping. The analytical capability of this platform is demonstrated in several applications, such as digital assay characterization, detection and assessment of DNA methylation heterogeneity, and detection of rare biomarkers at frequencies as low as 0.0005% target to background molecules.