A new class of cleavable fluorescent nucleotides: synthesis and optimization as reversible terminators for DNA sequencing by synthesis.
- Turcatti G Manteia Predictive Medicine S.A., Zone Industrielle, Coinsins, CH-1267, Switzerland. gerardo.turcatti@epfl.ch
- Romieu A
- Fedurco M
- Tairi AP
- 2008-02-12
Published in:
- Nucleic acids research. - 2008
Deoxyribonucleotides
Fluorescent Dyes
Microscopy, Fluorescence
Organic Chemicals
Sequence Analysis, DNA
English
Fluorescent 2'-deoxynucleotides containing a protecting group at the 3'-O-position are reversible terminators enabling array-based DNA sequencing by synthesis (SBS) approaches. Herein, we describe the synthesis of a new family of 3'-OH unprotected cleavable fluorescent 2'-deoxynucleotides and their evaluation as reversible terminators for high-throughput DNA SBS strategies. In this first version, all four modified nucleotides bearing a cleavable disulfide Alexa Fluor(R) 594 dye were assayed for their ability to act as a reversible stop for the incorporation of the next labeled base. Their use in SBS leaded to a signal-no signal output after successive addition of each labeled nucleotide during the sequencing process (binary read-out). Solid-phase immobilized synthetic DNA target sequences were used to optimize the method that has been applied to DNA polymerized colonies or clusters obtained by in situ solid-phase amplification of fragments of genomic DNA templates.
- Language
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- English
- Open access status
- gold
- Identifiers
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- DOI 10.1093/nar/gkn021
- PMID 18263613
- Persistent URL
- https://sonar.ch/global/documents/225486
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