ArticleNucleic acids research2003
Incorporation of reporter molecule-labeled nucleotides by DNA polymerases. I. Chemical synthesis of various reporter group-labeled 2'-deoxyribonucleoside-5'-triphosphates.
Article in Nucleic acids research, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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17 citing papers in PubMed, 71 citations in OpenAlex.
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- dUTPs conjugated with zwitterionic Cy3 or Cy5 fluorophore analogues are effective substrates for DNA amplification and labelling by Taq polymerase.Nucleic acids research · 2018Article
- Measurement of incorporation kinetics of non-fluorescent native nucleotides by DNA polymerases using fluorescence microscopy.Nucleic acids research · 2017Article
- Comparative Study of Novel Fluorescent Cyanine Nucleotides: Hybridization Analysis of Labeled PCR Products Using a Biochip.Journal of fluorescence · 2017Article
- Efficient enzymatic synthesis and dual-colour fluorescent labelling of DNA probes using long chain azido-dUTP and BCN dyes.Nucleic acids research · 2016Article
- The Toolbox for Modified Aptamers.Molecular biotechnology · 2016Review
- Structural basis for the synthesis of nucleobase modified DNA by Thermus aquaticus DNA polymerase.Proceedings of the National Academy of Sciences of the United States of America · 2010Article
- Photoinitiator nucleotide for quantifying nucleic Acid hybridization.Biomacromolecules · 2010Article
- High density labeling of polymerase chain reaction products with the fluorescent base analogue tCo.Analytical chemistry · 2009Article
- Highly efficient incorporation of the fluorescent nucleotide analogs tC and tCO by Klenow fragment.Nucleic acids research · 2009Article
- A new class of cleavable fluorescent nucleotides: synthesis and optimization as reversible terminators for DNA sequencing by synthesis.Nucleic acids research · 2008Article
- Systematic characterization of 2'-deoxynucleoside- 5'-triphosphate analogs as substrates for DNA polymerases by polymerase chain reaction and kinetic studies on enzymatic production of modified DNA.Nucleic acids research · 2006Article
- Real-time detection of polymerase activity using supercritical angle fluorescence.Journal of fluorescence · 2004Article
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6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Fluorescent-labeled DNA is generated through enzymatic incorporation of fluorophore-linked 2'-deoxyribonucleoside-5'-triphosphates (dNTPs) by DNA polymerases. We describe the synthesis of a variety of dye-labeled dNTPs. Amino-linker-modified 5'-triphosphates of all four naturally occurring nucleobases were used as precursors. Commercially available dyes were coupled to the amino function of the side chain. In addition, we attached novel fluorophore derivatives. The labeled products were obtained in at least 96% purity after HPLC purification. Enzymatic incorporation into DNA and subsequent extension of the modified DNA chain were studied. Vent(R) exo- DNA polymerase and a defined template-primer system were used to analyze each dye-labeled dNTP derivative. Our data suggest that the incorporation efficiency depends on the selected dye, the nucleobase or a combination of both.
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