Further we have used genome sequence independent microsatellites to identify global differences in the genomes of 93 cancer, cancer-free and high risk patient cell line samples [23]. This paper describes a larger high density oligonucleotide microarray with 370,000 elements, called Universal Bio-signature Detection Array (UBDA), designed by our laboratory and commercially produced by Roche-Nimblegen (Madison, WI) using light-directed photolithography [16, 24]. The platform design which consists mainly of probes, that are tailored to be genome independent, is mathematically derived and therefore unbiased (Additional file 1, Table S1). This strategy exploits the unique signature of a sample
in the form of Selleckchem Screening Library a pattern generated from hybridization of any unknown genome (DNA or cDNA) to a very high-density species-independent oligonucleotide microarray. Brucella species and several other pathogens were used as examples to demonstrate this forensics technology platform. Hybridization patterns are unique to a genome, and potentially to different isolates or a mixture of organisms. These techniques may be especially useful in evaluating and differentiating species whose genome has not yet been sequenced. Results UBDA array
sensitivity and specificity BGB324 nmr of probe hybridization DNA microarrays using oligonucleotides are widely used in biological research and are usually sequence specific. Two primary types of parameters are required to evaluate the robustness and sensitivity of DNA microarray experiments- labelling and hybridization [16]. Sensitivity of a given array platform is often defined as the minimum signal detected by the array scanning system [25]. In our case we have used labelling controls, where specified DNA molecules (CHIR98014 supplier 70-mer oligonucleotides) are
spiked into experimental human genomic DNA samples prior to fluorescent labelling. A set of six synthetic 70-mer oligonucleotides oxyclozanide (Additional file 2, Table S2) was designed to be spiked into each labelling reaction and hybridized to a constellation of 361 probes that were replicated five times on the array. We compared signal intensity values from control probes on the array hybridized with human genomic DNA and 70-mer oligonucleotides spiked into a separate sample of human genomic DNA. Each spike-in concentration was added on an individual array. We measured sensitivity of the array as a decrease in the correlation coefficient R2 value in the signal intensity from human genomic DNA spiked with 70-mer oligonucleotides when compared to the unspiked human genomic DNA sample. The sensitivity of the UBDA was examined by the addition of 70-mer synthetic oligonucleotides to the labeling reaction of human genomic DNA sample (Cy-3 label). Spike-in control synthetic 70-mer oligonucleotides were added at varying concentrations; 4.