Terminal phosphate-labeled nucleotides with improved substrate properties for homogeneous nucleic acid assays.
Nucleotides with a dye attached to the terminal phosphate with four or more phosphates (tetra- or pentaphosphates) are superior substrates than the corresponding triphosphates for DNA and RNA polymerases. When fluorogenic dyes are directly attached to the terminal phosphate, they can be released by the action of polymerase and alkaline phosphatase. The released dye changes color and fluorescence properties. The fluorescent signal can also be amplified by using multiple labeled nucleotides to detect small amounts of template. We have explored the utility of these nucleotides in a variety of applications including homogeneous SNP detection methods, DNA sequencing, and quantitation of PCR and RCA.
Sood
Anup
A
GE Healthcare, 800 Centennial Avenue, Piscataway, New Jersey 08855, USA.
Kumar
Shiv
S
Nampalli
Satyam
S
Nelson
John R
JR
Macklin
John
J
Fuller
Carl W
CW
D004247:DNA / Q000032:analysis* / Q000737:chemistry; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D009711:Nucleotides / Q000138:chemical synthesis / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D012313:RNA / Q000032:analysis* / Q000737:chemistry
D005456:Fluorescent Dyes; D009711:Nucleotides; D012313:RNA; D004247:DNA