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High-throughput approach for detection of DNA bending and flexibility based on cyclization.

Authors
  • Zhang, Yongli
  • Crothers, Donald M
Type
Published Article
Journal
Proceedings of the National Academy of Sciences
Publisher
Proceedings of the National Academy of Sciences
Publication Date
Mar 18, 2003
Volume
100
Issue
6
Pages
3161–3166
Identifiers
PMID: 12629220
Source
Medline
License
Unknown

Abstract

We have developed a high-throughput approach to the labor-intensive problems of DNA cyclization, which we use to characterize DNA curvature and mechanical properties. The method includes a combinatorial approach to make the DNA constructs needed and automated real-time measurement of the kinetics using fluorescence. We validated the approach and investigated the flexibility of two kinds of nicked DNA and AT dinucleotide repeats. We found that, although the nicks hardly alter the bending flexibility, they significantly increase the torsional flexibility, and that the AT repeat has 28% (+/-12%) lower bending rigidity than a generic DNA sequence.

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