Simple method of DNA stretching on glass substrate for fluorescence imaging and spectroscopy

Abstract

We demonstrate a simple method of stretching DNA to its full length, suitable for optical imaging and atomic force microscopy (AFM). Two competing forces on the DNA molecules, which are the electrostatic attraction between positively charged dye molecules (YOYO-1) intercalated into DNA and the negatively charged surface of glass substrate, and the centrifugal force of the rotating substrate, are mainly responsible for the effective stretching and the dispersion of single strands of DNA. The density of stretched DNA molecules could be controlled by the concentration of the dye-stained DNA solution. Stretching of single DNA molecules was confirmed by AFM imaging and the photoluminescence spectra of single DNA molecule stained with YOYO-1 were obtained, suggesting that our method is useful for spectroscopic analysis of DNA at the single molecule level. © 2014 Society of Photo-Optical Instrumentation Engineers.