Construction of branched DNA for SNP determination on glass-chip using photochemical ligation

Abstract

Single nucleotide polymorphisms (SNPs) are the most frequent form of sequence variation among individuals. Besides, sequence specific DNA detection has been an issue for its application in the diagnosis of pathogenic and genetic diseases. Many detection techniques that rely upon target hybridization with radioactive, fluorescent, chemiluminescent, and other types of label probes have been developed so far. Chip based detection for analysis of biomedical features like SNPs is not widely used in routine clinical settings, because it needs lot of improvement in terms of reproducibility and fidelity. Here, we have shown SNP typing on glass chip using KRAS oncogene sequence as a model. We have also shown that this SNP typing can be multiplexed by the further optimization of the photoligation protocol for high throughput detection that may lead to determine the sequence selectivity, less restricted reaction conditions and high efficiency.