Application of equilibrium models of solution hybridization to microarray design and analysis.

TitleApplication of equilibrium models of solution hybridization to microarray design and analysis.
Publication TypeJournal Article
Year of Publication2010
AuthorsGharaibeh RZ, Newton JM, Weller JW, Gibas CJ
JournalPLoS One
Volume5
Issue6
Paginatione11048
Date Published2010 Jun 10
ISSN1932-6203
KeywordsDNA Probes, Nucleic Acid Hybridization, Oligonucleotide Array Sequence Analysis
Abstract

<p><b>BACKGROUND: </b>The probe percent bound value, calculated using multi-state equilibrium models of solution hybridization, is shown to be useful in understanding the hybridization behavior of microarray probes having 50 nucleotides, with and without mismatches. These longer oligonucleotides are in widespread use on microarrays, but there are few controlled studies of their interactions with mismatched targets compared to 25-mer based platforms.</p><p><b>PRINCIPAL FINDINGS: </b>50-mer oligonucleotides with centrally placed single, double and triple mismatches were spotted on an array. Over a range of target concentrations it was possible to discriminate binding to perfect matches and mismatches, and the type of mismatch could be predicted accurately in the concentration midrange (100 pM to 200 pM) using solution hybridization modeling methods. These results have implications for microarray design, optimization and analysis methods.</p><p><b>CONCLUSIONS: </b>Our results highlight the importance of incorporating biophysical factors in both the design and the analysis of microarrays. Use of the probe "percent bound" value predicted by equilibrium models of hybridization is confirmed to be important for predicting and interpreting the behavior of long oligonucleotide arrays, as has been shown for short oligonucleotide arrays.</p>

DOI10.1371/journal.pone.0011048
Alternate JournalPLoS ONE
PubMed ID20548788
PubMed Central IDPMC2883574
Grant ListR01 GM072619 / GM / NIGMS NIH HHS / United States
1R01GM072619-01 / GM / NIGMS NIH HHS / United States