PMID: 8972863Dec 1, 1996Paper

Real-time imaging of the reorientation mechanisms of YOYO-labelled DNA molecules during 90 degrees and 120 degrees pulsed field gel electrophoresis

Nucleic Acids Research
S GurrieriC Bustamante

Abstract

Pulsed field gel electrophoresis (PFGE) techniques have been developed to overcome the limitations of conventional electrophoresis and to increase the separation to DNA chromosomes of few megabase pairs in size. Despite of the large success of these techniques, the various separation protocols employed for PFGE experiments have been determined empirically. However, a deep understanding of the molecular mechanisms of motion responsible for DNA separation becomes necessary for the rational optimization of these techniques. This paper shows the first clear observations of individual molecules of DNA during the reorientation process in 90 degrees PFGE and 120 degrees PFGE. Real-time visualization of the DNA dynamics during PFGE was possible with the use of an epi-illumination fluorescence microscope specifically equipped to run these experiments and by staining the DNA with YOYO-1 (1,1'-(4,4,7,7-tetramethyl-4,7-diazaundecamethylene)-bis-4-[3-meth yl -2,3-dihydro-(benzo-1,3-oxazole)-2-methyl-idene]-quinolinium tetraiodide). This dye forms a very stable, highly fluorescent complex with double-stranded DNA and dramatically improves the quality of the DNA images. The results of computer simulations used to reproduce the molecular mecha...Continue Reading

References

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Citations

Jan 14, 2000·Electrophoresis·S KatsuraA Mizuno
Dec 1, 2002·Journal of Biological Physics·A Mizuno, S Katsura
Feb 17, 2011·The Journal of Chemical Physics·Renat N Khaliullin, Jay D Schieber
Jan 20, 1999·Proceedings of the National Academy of Sciences of the United States of America·S GurrieriC Bustamante
Sep 14, 2013·Chemical Society Reviews·David R AlmassianWilliam M Nelson
Dec 23, 2008·Journal of Chromatography. a·Nancy C Stellwagen, Earle Stellwagen
Sep 8, 2004·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Jeanne L UyGer van den Engh
Mar 17, 2010·Angewandte Chemie·Neda NazemifardD Jed Harrison
May 10, 2020·The Analyst·Xuelin JinKyubong Jo
Oct 26, 2002·Physical Review Letters·Dieter Braun, Albert Libchaber

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