Abstract

DNA microarrays constitute an in vitro example system of a highly crowded molecular recognition environment. Although they are widely applied in many biological applications, some of the basic mechanisms of the hybridization processes of DNA remain poorly understood. On a microarray, cross-hybridization arises from similarities of sequences that may introduce errors during the transmission of information. Experimentally, we determine an appropriate distance, called minimum Hamming distance, in which the sequences of a set differ. By applying an algorithm based on a graph-theoretical method, we find large orthogonal sets of sequences that are sufficiently different not to exhibit any cross-hybridization. To create such a set, we first derive an analytical solution for the number of sequences that include at least four guanines in a row for a given sequence length and eliminate them from the list of candidate sequences. We experimentally confirm the orthogonality of the largest possible set with a size of 23 for the length of 7. We anticipate our work to be a starting point toward the study of signal propagation in highly competitive environments, besides its obvious application in DNA high throughput experiments.

Highlights

  • Molecular recognition in the crowded environment of DNA microarrays plays an important role in processing information

  • Because the longest sequences studied with our algorithm are 7-mers, we design a microarray consisting of oligonucleotides of length 7

  • All intensities are normalized relative to the average perfectly matching target (PM) intensity on the microarray

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Summary

Introduction

Molecular recognition in the crowded environment of DNA microarrays plays an important role in processing information. For DNA, specific-binding of two single strands, that is the formation of a stable double helix, occurs only if the bases A and T as well as C and G pair along the sequence. DNA microarrays are a widely used platform that, besides many applications in medicine and biology, enables the study of the fundamentals of DNA hybridization.[2−10] These microarrays consist of single-stranded DNA oligonucleotides immobilized on a surface (probes). If these probes are exposed to a bulk mixture of fluorescently labeled target sequences, only complementary targets are expected to hybridize. On a DNA microarray with complex target mixtures, imperfect recognition introduces noise and makes results difficult to interpret

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