Analytical and Bioanalytical Chemistry

, Volume 408, Issue 23, pp 6453–6459

Direct and precise length measurement of single, stretched DNA fragments by dynamic molecular combing and STED nanoscopy

  • Namdoo Kim
  • Hyung Jun Kim
  • Younggyu Kim
  • Kyung Suk Min
  • Seong Keun Kim
Research Paper

Abstract

A combination of DNA stretching method and super-resolution nanoscopy allows an accurate and precise measurement of the length of DNA fragments ranging widely in size from 117 to 23,130 bp. BstEII- and HindIII-treated λDNA fragments were stained with an intercalating dye and then linearly stretched on a coverslip by dynamic molecular combing. The image of individual DNA fragments was obtained by stimulated emission depletion nanoscopy. For DNA fragments longer than ∼1000 bp, the measured lengths of DNA fragments were consistently within ∼0.5 to 1.0 % of the reference values, raising the possibility of this method in a wide range of applications including facile detection for copy number variations and trinucleotide repeat disorder.

Keywords

DNA Dynamic molecular combing Length measurement STED nanoscopy 

Supplementary material

216_2016_9764_MOESM1_ESM.pdf (369 kb)
ESM 1(PDF 369 kb)

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Namdoo Kim
    • 1
  • Hyung Jun Kim
    • 1
  • Younggyu Kim
    • 1
    • 2
  • Kyung Suk Min
    • 3
  • Seong Keun Kim
    • 1
    • 3
  1. 1.Department of ChemistrySeoul National UniversitySeoulRepublic of Korea
  2. 2.LumiMac, Inc.SeoulKorea
  3. 3.Department of Biophysics and Chemical BiologySeoul National UniversitySeoulRepublic of Korea

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