ArticleProceedings of the National Academy of Sciences of the United States of America2005
Four-color DNA sequencing by synthesis on a chip using photocleavable fluorescent nucleotides.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed.
- Article
- Early monitoring-to-warning Internet of Things system for emerging infectious diseases via networking of light-triggered point-of-care testing devices.Exploration (Beijing, China) · 2023Article
- Challenges Related to the Use of Next-Generation Sequencing for the Optimization of Drug Therapy.Handbook of experimental pharmacology · 2023Article
- Enzymatic synthesis of hypermodified DNA polymers for sequence-specific display of four different hydrophobic groups.Nucleic acids research · 2020Article
- Long walk to genomics: History and current approaches to genome sequencing and assembly.Computational and structural biotechnology journal · 2020Review
- PyFrag 2019-Automating the exploration and analysis of reaction mechanisms.Journal of computational chemistry · 2019Article
- Structural Distortion of Cycloalkynes Influences Cycloaddition Rates both by Strain and Interaction Energies.Chemistry (Weinheim an der Bergstrasse, Germany) · 2019Article
- DNA sequencing at 40: past, present and future.Nature · 2017Review
- Molecular diagnostic testing by eyeGENE: analysis of patients with hereditary retinal dystrophy phenotypes involving central vision loss.Investigative ophthalmology & visual science · 2014Article
- Multicolor and erasable DNA photolithography.ACS nano · 2014Article
- The evolution of nanopore sequencing.Frontiers in genetics · 2014Review
- DNA branch migration reactions through photocontrollable toehold formation.Journal of the American Chemical Society · 2013Article
- Improved nucleotide selectivity and termination of 3'-OH unblocked reversible terminators by molecular tuning of 2-nitrobenzyl alkylated HOMedU triphosphates.Nucleic acids research · 2011Article
- 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
- An integrated system for DNA sequencing by synthesis using novel nucleotide analogues.Accounts of chemical research · 2010Article
- The challenges of sequencing by synthesis.Nature biotechnology · 2009Review
- 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
- A spatial dissection of the Arabidopsis floral transcriptome by MPSS.BMC plant biology · 2008Article
- A new class of cleavable fluorescent nucleotides: synthesis and optimization as reversible terminators for DNA sequencing by synthesis.Nucleic acids research · 2008Article
- 3'-O-modified nucleotides as reversible terminators for pyrosequencing.Proceedings of the National Academy of Sciences of the United States of America · 2007Article
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9 authors.
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Abstract
We report four-color DNA sequencing by synthesis (SBS) on a chip, using four photocleavable fluorescent nucleotide analogues (dGTP-PC-Bodipy-FL-510, dUTP-PC-R6G, dATP-PC-ROX, and dCTP-PC-Bodipy-650) (PC, photocleavable; Bodipy, 4,4-difluoro-4-bora-3alpha,4alpha-diaza-s-indacene; ROX, 6-carboxy-X-rhodamine; R6G, 6-carboxyrhodamine-6G). Each nucleotide analogue consists of a different fluorophore attached to the 5 position of the pyrimidines and the 7 position of the purines through a photocleavable 2-nitrobenzyl linker. After verifying that these nucleotides could be successfully incorporated into a growing DNA strand in a solution-phase polymerase reaction and the fluorophore could be cleaved using laser irradiation ( approximately 355 nm) in 10 sec, we then performed an SBS reaction on a chip that contains a self-priming DNA template covalently immobilized by using 1,3-dipolar azide-alkyne cycloaddition. The DNA template was produced by PCR, using an azido-labeled primer, and the self-priming moiety was attached to the immobilized DNA template by enzymatic ligation. Each cycle of SBS consists of the incorporation of the photocleavable fluorescent nucleotide into the DNA, detection of the fluorescent signal, and photocleavage of the fluorophore. The entire process was repeated to identify 12 continuous bases in the DNA template. These results demonstrate that photocleavable fluorescent nucleotide analogues can be incorporated accurately into a growing DNA strand during a polymerase reaction in solution and on a chip. Moreover, all four fluorophores can be detected and then efficiently cleaved using near-UV irradiation, thereby allowing continuous identification of the DNA template sequence. Optimization of the steps involved in this SBS approach will further increase the read-length.
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