Effect of DNA Concentration on Band Intensity and Resolution in Agarose Gel Electrophoresis

Year 2021, , 326 - 333, 30.12.2021

Mevlut Arslan Erdem Tezcan Haluk Camcı Murat Kemal Avcı

https://doi.org/10.52976/vansaglik.969547

Cited By: 2

Abstract

Objective: Agarose gel electrophoresis (AGE) is a widely used method for separating, identifying, and purifying nucleic acids. It has been used intensively in molecular researches and biotechnological applications. In AGE, band resolution and band quality are important parameters for researchers. Until now, many factors such as comb thickness, gel concentration, voltage and buffers have been reported to influence resolution and band quality in AGE. However, effect of DNA concentration on band resolution and intensity in AGE is unclear. Therefore, aim of the study was to investigate the effect of DNA concentration on AGE results in detail.
Materials and Methods: Different concentrations of DNA Marker and specific DNA fragments obtained by PCR were analyzed, but the loaded total DNA quantity was not changed. Furthermore, the effect of DNA concentration on AGE was also investigated by designed gel combs with different thicknesses and wideness.
Findings: It was shown that DNA concentration did not affect gel resolution, and the effect was not changed by comb thickness and wideness. Also, 2-fold dilution did not affect band intensities while 8-fold dilution significantly affected band intensities of all tested DNA fragments.
Conclusion: Concentrating samples, at least 8-fold, and using thin and narrow gel combs can be used for desired results in AGE.

Keywords

Agarose gel electrophoresis, DNA concentration, band quality, band intensity, gel resolution

Effect of DNA Concentration on Band Intensity and Resolution in Agarose Gel Elec-trophoresis

Abstract

Objective: Agarose gel electrophoresis (AGE) is a widely used method for separating, identifying, and purifying nucleic acids. It has been used intensively in molecular researches and biotechnological applications. In AGE, band resolution and band quality are important parameters for researchers. Until now, many factors such as comb thickness, gel concentration, voltage and buffers have been reported to influence resolution and band quality in AGE. However, effect of DNA concentration on band resolution and intensity in AGE is unclear. Therefore, aim of the study was to investigate the effect of DNA concentration on AGE results in detail.
Material and Method: Different concentrations of DNA Marker and specific DNA fragments obtained by PCR were analyzed, but the loaded total DNA quantity was not changed. Furthermore, the effect of DNA concentra-tion on AGE was also investigated by designed gel combs with different thicknesses and wideness.
Results: It was shown that DNA concentration did not affect gel resolution, and the effectwas not changed by comb thickness and wideness. Also, 2-fold dilution did not affect band intensities while 8-fold dilution signifi-cantly affected band intensities of all tested DNA fragments.
Conclusion: Concentrating samples, at least 8-fold, and using thin and narrow gel combs can be used for de-sired results in AGE.

Keywords

Agarose gel electrophoresis, DNA concentration, Band quality, Band intensity, Gel resolution

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