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A kinetically controlled, isothermal method for the detection of single nucleotide mismatches
Bioorganic & Medicinal Chemistry Letters ( IF 2.7 ) Pub Date : 2018-02-14 , DOI: 10.1016/j.bmcl.2018.02.024
Yoshiaki Masaki , Devon Cayer , Ryan McBride , M. Reza Ghadiri

We describe an isothermal, enzyme-free method to detect single nucleotide differences between oligonucleotides of close homology. The approach exploits kinetic differences in toe-hold-mediated, nucleic acid strand-displacement reactions to detect single nucleotide polymorphisms (SNPs) with essentially “digital” precision. The theoretical underpinning, experimental analyses, predictability, and accuracy of this new method are reported. We demonstrate detection of biologically relevant SNPs and single nucleotide differences in the let-7 family of microRNAs. The method is adaptable to microarray formats, as demonstrated with on-chip detection of SNP variants involved in susceptibility to the therapeutic agents abacavir, Herceptin, and simvastatin.



中文翻译:

动力学控制的等温方法,用于检测单核苷酸错配

我们描述了一种等温,无酶的方法来检测紧密同源的寡核苷酸之间的单核苷酸差异。该方法利用脚趾保持介导的核酸链置换反应中的动力学差异,以基本“数字”精度检测单核苷酸多态性(SNP)。报告了这种新方法的理论基础,实验分析,可预测性和准确性。我们证明了在let-7家族的microRNA的生物学相关的SNPs和单核苷酸差异的检测。该方法适用于微阵列格式,如芯片上检测涉及对治疗剂阿巴卡韦,赫赛汀和辛伐他汀易感性的SNP变体所证明的。

更新日期:2018-02-14
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