Spin-stretching of DNA and protein molecules for detection by fluorescence and atomic force microscopy.
We have developed a rapid and efficient way of stretching DNA and denatured protein molecules for detection by fluorescence microscopy and atomic force microscopy (AFM). In the described method, a viscous drag created by transient rotational flow stretches randomly coiled DNA molecules or denatured proteins. Stretching is achieved by dispensing a droplet of sample solution containing DNA or denatured protein on a MgCl2-soaked mica surface. We present fluorescent images of straightened lambdaDNA molecules and AFM images of stress-shared, reduced von Willebrand factor as well as straightened lambdaDNA. The described quick and reliable spin-stretching technique will find wide applications in the analysis of single biopolymer molecules.
Yokota
H
H
Department of Molecular, University of Washington, Seattle, Washington 98195, USA. hiroki@anatomy.iupui.edu
Sunwoo
J
J
Sarikaya
M
M
van den Engh
G
G
Aebersold
R
R
D004247:DNA / Q000032:analysis* / Q000737:chemistry; D018625:Microscopy, Atomic Force / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D011489:Protein Denaturation; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D014841:von Willebrand Factor / Q000032:analysis / Q000737:chemistry
D011506:Proteins; D014841:von Willebrand Factor; D004247:DNA