Adsorption models of hybridization and post-hybridization behaviour on oligonucleotide microarrays
Adsorption models of hybridization and post-hybridization behaviour on oligonucleotide microarrays
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DOI：
10.1088/0953-8984/18/23/024
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发表时间：
2006-06-14
影响因子：
2.7
通讯作者：
Wilson, Susan R.
中科院分区：
文献类型：
--
作者：
Burden, Conrad J.;Pittelkow, Yvonne;Wilson, Susan R.
对来自Affyssin Latin Square加标实验的数据的分析表明，寡核苷酸微阵列上特征的测量荧光强度通过双曲线响应函数与加标RNA靶浓度相关，该双曲线响应函数通常被鉴定为朗缪尔吸附等温线。此外，高加标浓度下的渐近信号对于失配特征几乎总是低于其伙伴完美匹配特征。我们调查了一些理论吸附模型的杂交在微阵列表面，并发现，在一般情况下，他们无法解释不同的饱和度响应的完美和不匹配的功能。另一方面，我们发现，一个简单而一致的解释，可以发现在一个模型中，平衡杂交后的双链体部分解离在杂交后的洗涤阶段。
Analysis of data from an Affymetrix Latin Square spike-in experiment indicates that measured fluorescence intensities of features on an oligonucleotide microarray are related to spike-in RNA target concentrations via a hyperbolic response function, generally identified as a Langmuir adsorption isotherm. Furthermore, the asymptotic signal at high spike-in concentrations is almost invariably lower for a mismatch feature than for its partner perfect match feature. We survey a number of theoretical adsorption models of hybridization at the microarray surface and find that in general they are unable to explain the differing saturation responses of perfect and mismatch features. On the other hand, we find that a simple and consistent explanation can be found in a model in which equilibrium hybridization is followed by partial dissociation of duplexes during the post-hybridization washing phase.