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.
CITATION STYLE
Kim, N., Kim, H. J., Kim, Y., Min, K. S., & Kim, S. K. (2016). Direct and precise length measurement of single, stretched DNA fragments by dynamic molecular combing and STED nanoscopy. Analytical and Bioanalytical Chemistry, 408(23), 6453–6459. https://doi.org/10.1007/s00216-016-9764-9
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