Hybridization isotherms of DNA microarrays and the quantification of mutation studies

Clinical Chemistry
A HalperinE B Zhulina

Abstract

Diagnostic DNA arrays for detection of point mutations as markers for cancer usually function in the presence of a large excess of wild-type DNA. This excess can give rise to false positives as a result of competitive hybridization of the wild-type target at the mutation spot. Analysis of the DNA array data is typically qualitative, aimed at establishing the presence or absence of a particular point mutation. Our theoretical approach yields methods for quantifying the analysis to obtain the ratio of concentrations of mutated and wild-type DNA. The theory is formulated in terms of the hybridization isotherms relating the hybridization fraction at the spot to the composition of the sample solutions at thermodynamic equilibrium. It focuses on samples containing an excess of single-stranded DNA and on DNA arrays with a low surface density of probes. The hybridization equilibrium constants can be obtained by the nearest-neighbor method. Two approaches allow acquisition of quantitative results from the DNA array data. In one, the signal of the mutation spot is compared with that of the wild-type spot. The implementation requires knowledge of the saturation intensity of the two spots. The second approach requires comparison of the int...Continue Reading

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Citations

Jun 29, 2006·Journal of the American Chemical Society·Gang ShenRastislav Levicky
Jan 12, 2013·Nucleic Acids Research·Andrew HarrisonAlexander E Pozhitkov
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May 28, 2009·The Journal of Physical Chemistry. B·Julia RazumovitchCorinne Vebert
May 6, 2017·ACS Omega·Mina Mohammadi-KambsAlbrecht Ott

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