Patent ID: 7179602
Filing Date: 2007-02-20
Classification: C12P,C12Q

Abstract:
1. A method of fluorescence-based cycle sequencing of a sample DNA, comprising, (a) preparing a reaction mixture containing: (b), dissociating the sample DNA to create single stranded templates, wherein said dissociation is achieved by heating the sample DNA to between about 92° C. and 95° C. for about 3 minutes; (c) annealing the primers to the primer sites, wherein said annealing is achieved at a temperature of between about 65° C. and 67° C. for about 30 seconds; (d) extending the annealed primers to generate a series of fluorescently-labeled dideoxynucleic acid fragments, wherein said primer extension is achieved at a temperature of between 75° C. and 78° C. for between about 3 to 4 minutes; (e) heating the reaction mixture to between about 92° C. and 95° C. in order to dissociate double stranded DNA; (f) repeating the steps c through e for a plurality of cycles; and (g) determining the nucleotide sequence of the sample DNA from the series of fluorescently-labeled dideoxynucleic acid fragments present in the reaction mixture.