ONE-TUBE POST-PCR FLUORESCENT LABELING OF DNA FRAGMENTS

A method for fluorescent postlabeling of PCR products has been develop ed. The method uses Klenow fragment of DNA polymerase I that exchanges the 3'-terminal residue of PCR-amplified DNA fragment for fluorescent nucleotides. All reactions, including PCR, are performed in one tube simply by successive addition of reagents. The products can be applied directly to fluorescence-based automated DNA sequencers without purif ication for either length determination in denaturing electrophoresis or mutation detection in SSCP electrophoresis.