ArticleProceedings of the National Academy of Sciences of the United States of America2007
3'-O-modified nucleotides as reversible terminators for pyrosequencing.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 49 citations in OpenAlex.
- Template-independent enzymatic synthesis of RNA oligonucleotides.Nature biotechnology · 2025Article
- Controlled enzymatic synthesis of oligonucleotides.Communications chemistry · 2024Review
- A new method to synthesize multiple gRNA libraries and functional mapping of mammalian H3K4me3 regions.Nucleic acids research · 2023Article
- Towards the controlled enzymatic synthesis of LNA containing oligonucleotides.Frontiers in chemistry · 2023Article
- Methods to improve the accuracy of next-generation sequencing.Frontiers in bioengineering and biotechnology · 2023Review
- Evaluation of the correctable decoding sequencing as a new powerful strategy for DNA sequencing.Life science alliance · 2022Article
- Large-ScaleFrontiers in bioengineering and biotechnology · 2021Review
- Mini review: Enzyme-based DNA synthesis and selective retrieval for data storage.Computational and structural biotechnology journal · 2021Review
- De novo DNA synthesis using polymerase-nucleotide conjugates.Nature biotechnology · 2018Article
- Mass-spectrometry analysis of modifications at DNA termini induced by DNA polymerases.Scientific reports · 2017Article
- The evolution of nanopore sequencing.Frontiers in genetics · 2014Review
- DNA polymerases drive DNA sequencing-by-synthesis technologies: both past and present.Frontiers in microbiology · 2014Review
- DNA sequencing by synthesis using 3'-RSC advances · 2014Article
- The history and advances of reversible terminators used in new generations of sequencing technology.Genomics, proteomics & bioinformatics · 2013Review
- DNA and RNA analyses in detection of genetic predisposition to cancer.Hereditary cancer in clinical practice · 2012Article
- Synthetic evolving systems that implement a user-specified genetic code of arbitrary design.Chemistry & biology · 2012Article
- Rapid incorporation kinetics and improved fidelity of a novel class of 3'-OH unblocked reversible terminators.Nucleic acids research · 2012Article
- An integrated system for DNA sequencing by synthesis using novel nucleotide analogues.Accounts of chemical research · 2010Article
- Reconstructed evolutionary adaptive paths give polymerases accepting reversible terminators for sequencing and SNP detection.Proceedings of the National Academy of Sciences of the United States of America · 2010Article
- Four-color DNA sequencing with 3'-O-modified nucleotide reversible terminators and chemically cleavable fluorescent dideoxynucleotides.Proceedings of the National Academy of Sciences of the United States of America · 2008Article
Corrections and comments
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Authors and funding
11 authors at 1 institution in 1 country.
Funding
Abstract
Pyrosequencing is a method used to sequence DNA by detecting the pyrophosphate (PPi) group that is generated when a nucleotide is incorporated into the growing DNA strand in polymerase reaction. However, this method has an inherent difficulty in accurately deciphering the homopolymeric regions of the DNA templates. We report here the development of a method to solve this problem by using nucleotide reversible terminators. These nucleotide analogues are modified with a reversible chemical moiety capping the 3'-OH group to temporarily terminate the polymerase reaction. In this way, only one nucleotide is incorporated into the growing DNA strand even in homopolymeric regions. After detection of the PPi for sequence determination, the 3'-OH of the primer extension products is regenerated through different deprotection methods. Using an allyl or a 2-nitrobenzyl group as the reversible moiety to cap the 3'-OH of the four nucleotides, we have synthesized two sets of 3'-O-modified nucleotides, 3'-O-allyl-dNTPs and 3'-O-(2-nitrobenzyl)-dNTPs as reversible terminators for pyrosequencing. The capping moiety on the 3'-OH of the DNA extension product is efficiently removed after PPi detection by either a chemical method or photolysis. To sequence DNA, templates containing homopolymeric regions are immobilized on Sepharose beads, and then extension-signal detection-deprotection cycles are conducted by using the nucleotide reversible terminators on the DNA beads to unambiguously decipher the sequence of DNA templates. Our results establish that this reversible-terminator-pyrosequencing approach can be potentially developed into a powerful methodology to accurately determine DNA sequences.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.