Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (453K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Capaldi, S.
Right arrow Articles by Jayasena, S. D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Capaldi, S.
Right arrow Articles by Jayasena, S. D.
Related Collections
Right arrow DNA characterisation
Right arrow Genomics
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2000, Vol. 28, No. 7 E21-e21
© 2000 Oxford University Press

Signal amplification through nucleotide extension and excision on a dendritic DNA platform

Stephen Capaldi, Robert C. Getts1 and Sumedha D. Jayasena*

NeXstar Pharmaceuticals Inc., 2860 Wilderness Place, Boulder, CO 80301, USA and 1Genisphere Inc., 4170 City Avenue, Philadelphia, PA 19131, USA

Techniques that provide strong signal amplification are useful in diagnostic applications, especially in detecting low concentrations of non-amplifiable target molecules. A versatile and strong signal amplification method based on activities of a DNA polymerase to generate high concentrations of pyrophosphate (PPi) is described. The generation of PPi is catalyzed by nucleotide extension and excision activities of a DNA polymerase on an oligonucleotide cassette. The signal is generated upon enzymatic conversion of PPi to ATP and ATP levels subsequently detected with firefly luciferase. Bioluminesence produced by an oligonucleotide cassette consisting of just two polymerase reaction sites is sufficient to detect them at low attomole levels. The attachment of a large number of these oligonucleotide cassettes to DNA dendrimers enabled the detection of such poly­valent substrate molecules at low zeptomole (10–21 mol) concentrations. The extent of signal amplification obtained with dendrimer substrates is comparable to exponential target amplifications provided by nucleic acid amplification methods. The attachment of such PPi-generating dendritic DNA platforms to ligands that mediate target recognition would potentially permit detection of extremely low concentrations of analytes in diagnostic assays.

* To whom correspondence should be addressed at: 5875 North Orchard Creek Circle, Boulder, CO 80301, USA. Tel: +1 303 581 9414; Email: sumedhaj@yahoo.com


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Nucleic Acids ResHome page
G.-h. Zhou, M. Kamahori, K. Okano, G. Chuan, K. Harada, and H. Kambara
Quantitative detection of single nucleotide polymorphisms for a pooled sample by a bioluminometric assay coupled with modified primer extension reactions (BAMPER)
Nucleic Acids Res., October 1, 2001; 29(19): e93 - e93.
[Abstract] [Full Text] [PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.