Skip Navigation

Nucleic Acids Research 2006 34(11):e81; doi:10.1093/nar/gkl261
This Article
Right arrow Full Text Freely available
Right arrow Print PDF (111K) Freely available
Right arrow Screen PDF (117K) 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 arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Yi, J.
Right arrow Articles by Zhang, D. Y.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Yi, J.
Right arrow Articles by Zhang, D. Y.
Related Collections
Right arrow Nucleic acid amplification
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Published online 5 July 2006

© 2006 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commerical use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

Molecular Zipper: a fluorescent probe for real-time isothermal DNA amplification

Jizu Yi, Wandi Zhang and David Y. Zhang*

Department of Pathology, Mount Sinai School of Medicine, New York University One Gustave L. Levy Place, New York, NY 10029, USA

*To whom correspondence should be addressed at Department of Pathology, Mount Sinai School of Medicine, New York University, One Gustave L. Levy Place, New York, NY 10029, USA. Tel: +1 212 659 8173; Fax: +1 212 427 2082; Email: david.zhang{at}mssm.edu

Received October 17, 2005. Revised November 21, 2005. Accepted March 31, 2006.

Rolling-circle amplification (RCA) and ramification amplification (RAM, also known as hyperbranched RCA) are isothermal nucleic acid amplification technologies that have gained a great application in in situ signal amplification, DNA and protein microarray assays, single nucleotide polymorphism detection, as well as clinical diagnosis. Real-time detection of RCA or RAM products has been a challenge because of most real-time detection systems, including Taqman and Molecular Beacon, are designed for thermal cycling-based DNA amplification technology. In the present study, we describe a novel fluorescent probe construct, termed molecular zipper, which is specially designed for quantifying target DNA by real-time monitoring RAM reactions. Our results showed that the molecular zipper has very low background fluorescence due to the strong interaction between two strands. Once it is incorporated into the RAM products its double strand region is opened by displacement, therefore, its fluorophore releases a fluorescent signal. Applying the molecular zipper in RAM assay, we were able to detect as few as 10 molecules within 90 min reaction. A linear relationship was observed between initial input of targets and threshold time (R2 = 0.985). These results indicate that molecular zipper can be applied to real-time monitoring and qualification of RAM reaction, implying an amenable method for automatic RAM-based diagnostic assays.

Present address: Jizu Yi, BD-Diagnostics, One Becton Drive, Franklin Lakes, NJ 07417, USA


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
 RNAHome page
B. Yao, J. Li, H. Huang, C. Sun, Z. Wang, Y. Fan, Q. Chang, S. Li, and J. Xi
Quantitative analysis of zeptomole microRNAs based on isothermal ramification amplification
RNA, September 1, 2009; 15(9): 1787 - 1794.
[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.