Flemish Institute for Technological Research (VITO), Boeretang 200, B-2400 Mol, Belgium.
Biosens Bioelectron. 2010 Dec 15;26(4):1692-5. doi: 10.1016/j.bios.2010.07.039. Epub 2010 Aug 16.
We consider mixtures of two DNA sequences t and t' differing by a single nucleotide, which are analyzed by an Agilent custom DNA microarray. In particular we focus on the case in which t, the "wild type", is predominantly abundant and t' the "mutant" is at very low concentrations compared to t. We show that by using appropriately designed arrays it is possible to accurately quantify the presence of t' even at low relative concentrations (≈1%). The detection method is based on thermodynamic models of DNA hybridisation and on the analysis of a large number of hybridisation intensities from probes containing one or two mismatches with respect to t and t'.
我们考虑两种 DNA 序列 t 和 t' 的混合物,它们仅在一个核苷酸处存在差异,并通过安捷伦定制 DNA 微阵列进行分析。特别地,我们关注的情况是,与 t(“野生型”)相比,t'(“突变型”)的丰度非常低。我们表明,通过使用适当设计的阵列,即使在相对低的浓度(≈1%)下,也可以准确地定量存在 t'。检测方法基于 DNA 杂交的热力学模型,以及对包含一个或两个与 t 和 t' 错配的探针的大量杂交强度的分析。