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

Guru P. Neupane; Krishna P. Dhakal; Hyunsoo Lee; Martin Guthold; Vincent S. Joseph; Jong-Dal Hong; Jeongyong Kim

doi:10.1117/12.2018692

20 May 2013 Simple method of DNA stretching on glass substrate for fluorescence image and spectroscopy

Guru P. Neupane, Krishna P. Dhakal, Hyunsoo Lee, Martin Guthold, Vincent S. Joseph, Jong-Dal Hong, Jeongyong Kim

Proceedings Volume 8879, Nano-Bio Sensing, Imaging, and Spectroscopy; 88790J (2013) https://doi.org/10.1117/12.2018692
Event: Nano-Bio Sensing, Imaging and Spectroscopy, 2013, Jeju, Korea, Republic of

Abstract

Study of biological molecule DNA has contributed to developing many breaking thoughts and wide applications in multidisciplinary fields, such as genomic, medical, sensing and forensic fields. Stretching of DNA molecules is an important supportive tool for AFM or spectroscopic studies of DNA in a single molecular level. In this article, we established a simple method of DNA stretching (to its full length) that occurred on a rotating negatively-charged surface of glass substrate. The isolation of a single DNA molecule was attained by the two competitive forces on DNA molecules, that is, the electrostatic attraction developed between the positively charged YOYO-1 stained DNA and the negatively charged substrate, and the centrifugal force of the rotating substrate, which separates the DNA aggregates into the single molecule. Density of stretched DNA molecules was controlled by selecting the specific parameters such as spinning time and rates, loading volume of DNA-dye complex solution etc. The atomic force microscopy image exhibited a single DNA molecule on the negatively-charged substrate in an isolated state. Further, the photoluminescence spectra of a single DNA molecule stained with YOYO-1 were achieved using the method developed in the present study, which is strongly believed to effectively support the spectroscopic analysis of DNA in a single molecular level.

Citation Download Citation

Guru P. Neupane, Krishna P. Dhakal, Hyunsoo Lee, Martin Guthold, Vincent S. Joseph, Jong-Dal Hong, and Jeongyong Kim "Simple method of DNA stretching on glass substrate for fluorescence image and spectroscopy", Proc. SPIE 8879, Nano-Bio Sensing, Imaging, and Spectroscopy, 88790J (20 May 2013); https://doi.org/10.1117/12.2018692

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