title: Direct and precise length measurement of single, stretched DNA fragments by dynamic molecular combing and STED nanoscopy.
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.
authors.affiliation: Department of Chemistry, Seoul National University, Gwanak-gu, Seoul, 151-747, Republic of Korea.
authors.forename: Namdoo
authors.initials: N
authors.lastname: Kim
authors.affiliation: Department of Chemistry, Seoul National University, Gwanak-gu, Seoul, 151-747, Republic of Korea.
authors.forename: Hyung Jun
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Department of Chemistry, Seoul National University, Gwanak-gu, Seoul, 151-747, Republic of Korea.|LumiMac, Inc., 4, Dongnam-ro 2-gil, B1, Songpa-gu, Seoul, Korea.
authors.forename: Younggyu
authors.initials: Y
authors.lastname: Kim
authors.affiliation: Department of Biophysics and Chemical Biology, Seoul National University, Gwanak-gu, Seoul, 151-747, Republic of Korea.
authors.forename: Kyung Suk
authors.initials: KS
authors.lastname: Min
authors.affiliation: Department of Chemistry, Seoul National University, Gwanak-gu, Seoul, 151-747, Republic of Korea. seongkim@snu.ac.kr.|Department of Biophysics and Chemical Biology, Seoul National University, Gwanak-gu, Seoul, 151-747, Republic of Korea. seongkim@snu.ac.kr.
authors.forename: Seong Keun
authors.initials: SK
authors.lastname: Kim
mesh_terms: D004247:DNA / Q000032:analysis*; D004867:Equipment Design; D018613:Microscopy, Confocal / Q000295:instrumentation* / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods; D056004:Molecular Dynamics Simulation; D036103:Nanotechnology / Q000295:instrumentation* / Q000379:methods
chemical_list: D004247:DNA
keywords: DNA; Dynamic molecular combing; Length measurement; STED nanoscopy