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

  • Author(s): Chan, Ting-Fung;
  • Ha, Connie;
  • Phong, Angie;
  • Cai, Dongmei;
  • Wan, Eunice;
  • Leung, Lucinda;
  • Kwok, Pui-Yan;
  • Xiao, Ming
  • et al.
Abstract

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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