Selectivity of Hybridization Controlled by the Density of Solid Phase Synthesized DNA Probes on Glass Substrates

摘要: Optical biochip design based on varying the density of immo- bilized single-stranded DNA (ssDNA) oligonucleotide probes was examined. A method immobilization developed to yield various densities probe molecules using photochemical activation surfaces and in situ solid phase synthesis for immobilization. High surface ssDNA (up 1 × 10 13 probes/cm 2 ) obtained method. The extent hybridization nucleic acids were determined confocal fluorescence microscopy. Selective targets associated with spinal muscular atrophy containing single nucleo- tide polymorphisms (SNP), their thermal denaturation profiles investigated examine sensitivity selectivity SNP detection. detection limit less than 16 pM at room temperature. Single base mismatch discrimination demonstrated control melt temp- erature by selection density, temperature differences 12- 15°C could be achieved determination. Importantly, results demonstrate that poor can result significant variability selectivity, as seen shifts up 5°C range investigated.