7-Deaza-2′-Deoxyadenosine-5′-Triphosphate as an Alternative Nucleotide for the Pyrosequencing Technology

A new adenosine nucleotide analog suitable for the Pyrosequencing method is presented. The new analog, 7-deaza-2′-deoxyadenosine-5′-triphosphate (c 7 dATP), has virtually the same low substrate specificity for luciferase as the currently used analog, 2′-deoxyadenosine-5′-O-(1-thiotriphosphate) (dATP...

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Bibliographic Details
Published in:Nucleosides, nucleotides & nucleic acids nucleotides & nucleic acids, 2004-01, Vol.23 (10), p.1583-1594
Main Authors: Eriksson †, Jonas, Gharizadeh †, Baback, Nourizad, Nader, Nyrén, Pål
Format: Article
Language:English
Subjects:
2′-deoxyadenosine-5′-O-(1-thiotriphosphate)
7-deaza-2 '-deoxyadenosine-5 '-triphosphate
Apyrase - antagonists & inhibitors
Base Sequence
Deoxyguanine Nucleotides - chemistry
DNA Primers
DNA sequencing
dna-polymerase
escherichia-coli
firefly luciferase
Luciferase
Luciferases - chemistry
Luciferases - metabolism
pyrophosphate
Pyrosequencing technology
Sequence Analysis, DNA - methods
Substrate Specificity
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Summary:A new adenosine nucleotide analog suitable for the Pyrosequencing method is presented. The new analog, 7-deaza-2′-deoxyadenosine-5′-triphosphate (c 7 dATP), has virtually the same low substrate specificity for luciferase as the currently used analog, 2′-deoxyadenosine-5′-O-(1-thiotriphosphate) (dATPαS). The inhibitory effect dATPαS displays on the nucleotide degrading activity of apyrase was reduced significantly by substituting the c 7 dATP for the dATPαS. Both analogs show high stability after long time storage at + 8°C. Furthermore, with the new nucleotide a read length of up to 100 bases was obtained for several templates from fungi, bacteria and viruses. † Equal contribution to this work.
ISSN:1525-7770
1532-2335
1532-2335
DOI:10.1081/NCN-200031402