One-tube post-PCR fluorescent labeling of DNA fragments.
A method for fluorescent postlabeling of PCR products has been developed. 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 purification for either length determination in denaturing electrophoresis or mutation detection in SSCP electrophoresis.
Inazuka
M
M
Division of Genome Analysis, Kyushu University, Fukuoka, Japan.
Tahira
T
T
Hayashi
K
K
D001331:Automation; D004247:DNA / Q000032:analysis*; D004256:DNA Polymerase I / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D019075:Electrophoresis, Capillary; D004591:Electrophoresis, Polyacrylamide Gel; D005452:Fluoresceins / Q000378:metabolism; D005453:Fluorescence; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D017421:Sequence Analysis / Q000379:methods
D017931:DNA Primers; D005452:Fluoresceins; D004247:DNA; D004256:DNA Polymerase I