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Nucleic Acids Research Advance Access published online on September 13, 2006
Nucleic Acids Research, doi:10.1093/nar/gkl593
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© 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-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

A simple DNA stretching method for fluorescence imaging of single DNA molecules

Ting-Fung Chan, Connie Ha, Angie Phong, Dongmei Cai, Eunice Wan, Lucinda Leung, Pui-Yan Kwok and Ming Xiao*

Cardiovascular Research Institute and Institute for Human Genetics University of California, San Francisco CA, USA

*To whom correspondence should be addressed. 513 Parnassus Avenue, HSW-901A, San Francisco, CA 94143, USA. Tel: +1 4155143876; Fax: +1 4154762956; Email: m.xiao{at}ucsf.edu

Received June 10, 2006. Revised July 28, 2006. Accepted July 31, 2006.

Stretching or aligning DNA molecules onto a surface by means of molecular combing techniques is one of the critical steps in single DNA molecule analysis. However, many of the current studies have focused on {lambda}-DNA, or other large DNA molecules. There are very few studies on stretching methodologies for DNA molecules generated via PCR (typically smaller than 20 kb). Here we describe a simple method of stretching DNA molecules up to 18 kb in size on a modified glass surface. The very low background fluorescence allows efficient detection of single fluorescent dye labels incorporated into the stretched DNA molecules.


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