Single nucleotide variation detection on 3D DNA microarray by ligation of two-terminal-modified universal probes

Exploiting the advantages of three-dimensional (3D) DNA microarray, we put forward a novel strategy termed "ligation of two-terminal-modified universal probes" for single nucleotide variation (SNV) detection on 3D microarray. By performing specific ligation reaction between the unmodified hybridization primer with 3' hydroxyl terminus and the universal probe with phosphorylated 5' terminus and fluorescently labeled 3' terminus, two point mutations (C3206T and A5301G) in the PCR products immobilized on the 3D polyacrylamide gel DNA microarray were accurately discriminated. This method can not only maintain the predominance of 3D DNA microarray as a platform with high through-put, but also can exert this predominance to the cases of detecting a small quantity of samples and multiple SNVs since four universal probes can be employed to detect all SNVs, therefore, it is more feasible for laboratory research and provide an effective tool for clinical diagnosis.