荧光团标记的引物提高了变性梯度凝胶电泳的灵敏度、通用性和标准化程度。
Fluorophore-labeled primers improve the sensitivity, versatility, and normalization of denaturing gradient gel electrophoresis.
作者信息
Neufeld Josh D, Mohn William W
机构信息
Department of Microbiology and Immunology, University of British Columbia, 300-6174 University Boulevard, Vancouver, British Columbia V6T 1Z3, Canada.
出版信息
Appl Environ Microbiol. 2005 Aug;71(8):4893-6. doi: 10.1128/AEM.71.8.4893-4896.2005.
Abstract
Denaturing gradient gel electrophoresis (DGGE) is widely used in microbial ecology. We tested the effect of fluorophore-labeled primers on DGGE band migration, sensitivity, and normalization. The fluorophores Cy5 and Cy3 did not visibly alter DGGE fingerprints; however, 6-carboxyfluorescein retarded band migration. Fluorophore modification improved the sensitivity of DGGE fingerprint detection and facilitated normalization of samples from multiple gels by the application of intralane standards.
摘要
变性梯度凝胶电泳（DGGE）在微生物生态学中被广泛应用。我们测试了荧光团标记引物对DGGE条带迁移、灵敏度和标准化的影响。荧光团Cy5和Cy3并未明显改变DGGE指纹图谱；然而，6-羧基荧光素会延迟条带迁移。荧光团修饰提高了DGGE指纹图谱检测的灵敏度，并通过应用泳道内标准物促进了来自多个凝胶的样品的标准化。
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