Okamoto Akimitsu, Tainaka Kazuki, Ochi Yuji, Kanatani Keiichiro, Saito Isao
Department of Synthetic Chemistry and Biological Chemistry, Faculty of Engineering, Kyoto University, Kyoto, 615-8510, Japan.
Mol Biosyst. 2006 Feb;2(2):122-7. doi: 10.1039/b515923g. Epub 2006 Jan 5.
We have developed a new concept involving a single-step homogeneous method for single-nucleotide polymorphism (SNP) typing. In this method, a probe containing base-discriminating fluorescent (BDF) bases is added to a sample solution. BDF base-containing DNA usually shows only a weak fluorescence, but emits a strong blue fluorescence when it recognizes a target base at a specific site in a hybridized strand. By utilizing this feature, a simple mix-and-read SNP typing assay was achieved without any tedious probe-designing or washing processes for exclusion of hybridization error or any addition of DNA-modifying enzymes. This is very different from conventional methods. We simultaneously analyzed a number of samples with ease, with a high accuracy, using our BDF assay.
我们开发了一种新概念，涉及用于单核苷酸多态性（SNP）分型的单步均相方法。在该方法中，将含有碱基区分荧光（BDF）碱基的探针添加到样品溶液中。含BDF碱基的DNA通常仅显示微弱荧光，但当它在杂交链的特定位点识别靶碱基时会发出强烈的蓝色荧光。利用这一特性，无需进行任何繁琐的探针设计或洗涤过程以排除杂交错误，也无需添加任何DNA修饰酶，即可实现简单的混合读取SNP分型检测。这与传统方法有很大不同。我们使用BDF检测轻松、高精度地同时分析了多个样品。
