Method to make fluorescent nucleotide photoproducts for dna sequencing and analysis

摘要

A method is described for making fluorescent photoproducts of nucleotides using certain solvents in degassed solutions under ultraviolet illumination. These photoproducts can be highly fluorescent (quantum yield QY on order of 50 %) in room temperature aqueous solutions and over a wide pH range, and have low bleaching probability on order of 10-4, with improvement anticipated with the use of triplet quenchers. These modified nucleotides, when bound and immobilized on an alumina surface, can be detected at the single molecule level. Their value lies in the significant enhancement in the detectability of nucleotides by fluorescence, and may offer an attractive set of fluorphores for application to base-at-a-time DNA sequencing.