Dna fragment sizing and sorting by laser-induced fluorescence

Details Dna fragment sizing and sorting by laser-induced fluorescence Dna fragment sizing and sorting by laser-induced fluorescence

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) G01N 15/14 (2006.01)

Patent

CA 2089423

DNA FRAGMENT SIZING AND SORTING BY LASER-INDUCED FLUORESCENCE ABSTRACT A method is provided for obtaining DNA fingerprints using high speed detection systems, such as flow cytometry to determine unique characteristics of DNA pieces from a selected sample. In one characterization the DNA piece is fragmented at preselected sites to produce a plurality of DNA fragments. The DNA piece or the resulting DNA fragments are treated with a dye effective to stain stoichiometrically the DNA fragments. The fluorescence from the dye in the stained fragments is then examined to generate an output functionally related to the number of nucleotides in each one of the DNA fragments. In one embodiment, the intensity of the fluorescence emissions from each fragment is directly proportional to the fragment length. Additional dyes can be bound to the DNA piece and DNA fragments to provide information additional to length information. Oligonucleotide specific dyes and/or hybridization probes can be bound to the DNA fragments to provide information on oligonucleotide distribution or probe hybridization to DNA fragments of different sizes.

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