A two-layer assay for single-nucleotide variants utilizing strand   displacement and selective digestion

doi:10.1016/j.bios.2016.03.070

CORC > 北京大学 > 化学与分子工程学院

	A two-layer assay for single-nucleotide variants utilizing strand displacement and selective digestion
	Yu, Yingjie ; Wu, Tongbo ; Johnson-Buck, Alexander ; Li, Lidan ; Su, Xin
刊名	BIOSENSORS & BIOELECTRONICS
	2016
关键词	Single nucleotide variant Toehold strand displacement Lambda exonuclease Fluorescent probe DNA HYBRIDIZATION MUTATIONS MEDICINE PLASMA
DOI	10.1016/j.bios.2016.03.070
英文摘要	Point mutations have emerged as prominent biomarkers for disease diagnosis, particularly in the case of cancer. Discovering single-nucleotide variants (SNVs) is also of great importance for the identification of single-nucleotide polymorphisms within the population. The competing requirements of thermodynamic stability and specificity in conventional nucleic acid hybridization probes make it challenging to achieve highly precise detection of point mutants. Here, we present a fluorescence-based assay for low abundance mutation detection based on toehold-mediated strand displacement and nuclease-mediated strand digestion that enables highly precise detection of point mutations. We demonstrate that this combined assay provides 50-1000-fold discrimination (mean value: 255) between all possible single nucleotide mutations and their corresponding wild-type sequence for a model DNA target. Using experiments and kinetic modeling, we investigate probe properties that obtain additive benefits from both strand displacement and nucleolytic digestion, thus providing guidance for the design of enzyme mediated nucleic acid assays in the future. (C) 2016 Elsevier B.V. All rights reserved.; National Natural Science Foundation of China [81273631]; subject promotion plan of Beijing University of Chemical Technology [ys12060026025]; SCI(E); EI; PubMed; ARTICLE; xinsu@mail.buct.edu.cn; 248-254; 82
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/434028]
专题	化学与分子工程学院
推荐引用方式 GB/T 7714	Yu, Yingjie,Wu, Tongbo,Johnson-Buck, Alexander,et al. A two-layer assay for single-nucleotide variants utilizing strand displacement and selective digestion[J]. BIOSENSORS & BIOELECTRONICS,2016.
APA	Yu, Yingjie,Wu, Tongbo,Johnson-Buck, Alexander,Li, Lidan,&Su, Xin.(2016).A two-layer assay for single-nucleotide variants utilizing strand displacement and selective digestion.BIOSENSORS & BIOELECTRONICS.
MLA	Yu, Yingjie,et al."A two-layer assay for single-nucleotide variants utilizing strand displacement and selective digestion".BIOSENSORS & BIOELECTRONICS (2016).