ChipcheckII—Predicting Binding Curves for Multiple Analyte Strands on Small DNA Microarrays

Abstract

Incomplete binding, saturation, and cross-hybridization between partially complementary strands complicate the parallel detection of nucleic acids via DNA microarrays. Treating the competing equilibria governing binding to microarrays requires computational tools. We have developed the web-based program ChipCheckII that calculates total hybridization matrices for target strands interacting with probes on small DNA microarrays. The program can be used to compute the extent of cross-hybridization and other phenomena affecting fidelity of detection based on sequences, quantities of strands, and hybridization conditions as inputs. Enthalpy and entropy of duplex formation are generated locally with UNAfold, including those for complexes that are partially matched. Simulated binding versus temperature curves for portions of a commercial genome chip demonstrate the extent to which cross-hybridization can complicate DNA detection. ChipCheckII is expected to aid nucleic acid chemists in developing high fidelity DNA microarrays.

Keywords:

• DNA
• microarray
• hybridization
• binding equilibria
• simulation

The authors wish to thank Dr. M. Bauer, Dr. U. Plutowski, and Prof. U. Steiner for helpful discussion. This work was supported by DFG (FOR 434, grant RI 1063/4).