Development of highly sensitive and low-cost DNA agarose gel electrophoresis detection systems, and evaluation of non-mutagenic and loading dye-type DNA-staining reagents
Autoři:
Ken Motohashi aff001
Působiště autorů:
Department of Frontier Life Sciences, Faculty of Life Sciences, Kyoto Sangyo University, Kamigamo Motoyama, Kita-ku, Kyoto, Japan
aff001; Center for Ecological Evolutionary Developmental Biology, Kyoto Sangyo University, Kamigamo Motoyama, Kita-Ku, Kyoto, Japan
aff002
Vyšlo v časopise:
PLoS ONE 14(9)
Kategorie:
Research Article
doi:
https://doi.org/10.1371/journal.pone.0222209
Souhrn
Highly sensitive and low-cost DNA agarose gel detection systems were developed using non-mutagenic and loading dye-type DNA-staining reagents. The DNA detection system that used Midori Green Direct and Safelook Load-Green, both with an optimum excitation wavelength at ~490 nm, could detect DNA-fragments at the same sensitivity to that of the UV (312 nm)-transilluminator system combined with ethidium bromide, after it was excited by a combination of cyan LED light and a shortpass filter (510 nm). The cyan LED system can be also applied to SYBR Safe that is widely used as a non-toxic dye for post-DNA-staining. Another DNA-detection system excited by black light was also developed. Black light used in this system had a peak emission at 360 nm and caused less damage to DNA due to lower energy of UV rays with longer wavelength when compared to those of short UV rays. Moreover, hardware costs of the black light system were ~$100, less than 1/10 of the commercially available UV (365 nm) transilluminator (>$1,000). EZ-Vision and Safelook Load-White can be used as non-mutagenic and loading dye-type DNA-staining reagents in this system. The black light system had a greater detection sensitivity for DNA fragments stained by EZ-Vision and Safelook Load-White compared with the commercially available imaging system using UV (365 nm) transilluminator.
Klíčová slova:
Biology and life sciences – Molecular biology – Molecular biology techniques – Molecular biology assays and analysis techniques – DNA filter assay – Genetics – DNA – DNA electrophoresis – Biochemistry – Nucleic acids – Research and analysis methods – Electrophoretic techniques – Gel electrophoresis – Agarose gel electrophoresis – Physical sciences – Physics – Electromagnetic radiation – Light – Artificial light – Ultraviolet radiation – Chemistry – Chemical compounds – Bromides
Zdroje
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