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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2019.1
- 수록면
- 158 - 164 (7page)
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초록· 키워드
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A We propose a novel quantitative assay for colorimetric loop-mediated isothermal amplification (LAMP) based on the shift of UV-vis spectral absorbance of Eriochrome black T (EBT) dye. The colorimetric LAMP assay provides on-and-off result of the LAMP reaction, not quantitative information. Since the quantification of the initial copy number of DNA is of importance for genetic diagnostics, we, for the first time, demonstrated how to quantify the DNA in the EBT mediated LAMP reaction. Due to the structral change of the EBT-Mg complexes, the color of the LAMP mixture is changed from violet to sky blue as the LAMP reaction proceeds. We noticed that the maxium absorption peak before and after the LAMP reaction is distinct, and accordingly shifted from 570 nm to 640 nm. We utilized the ratio between two particular absorbance (A640/A570) of EBT as an indicator for the LAMP process, and plotted it versus the reaction time. We used serially diluted DNA samples and produced quantitative calibration curves (the threshold time versus the initial copy number of DNA). Therefore, we could quantify the copy number of template DNAs in the EBT-mediated colorimetric LAMP reaction simply by measuring the two UV-vis absorption values at 570 nm and 640 nm.
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등록된 정보가 없습니다.
참고문헌 (21)
참고문헌 신청
Boon, N. / 2003 / Bioaugmentation as a tool to protect the structure and function of an activated-sludge microbial community against a 3-Chloroaniline shock load / Appl. Environ. Microbiol 69:1511~1520
Sharkey, F. H. / 2004 / Detection and quantification of gene expression in environmental bacteriology / Appl. Environ. Microbiol 70:3795~3806
Valones, M. A. A. / 2009 / Principles and applications of polymerase chain reaction in medical diagnostic fields: a review / Braz. J. Microbiol 40:1~11
Heid, C. A. / 1996 / Real time quantitative PCR / Genome Res 6:986~994
Giuffrida, M.C. / 2017 / Integration of isothermal amplification methods in microfluidic devices:Recent advances / Biosens. Bioelectron 90:174~186
Sharkey, F. H. / 2004 / Detection and quantification of gene expression in environmental bacteriology / Appl. Environ. Microbiol 70:3795~3806
Valones, M. A. A. / 2009 / Principles and applications of polymerase chain reaction in medical diagnostic fields: a review / Braz. J. Microbiol 40:1~11
Heid, C. A. / 1996 / Real time quantitative PCR / Genome Res 6:986~994
Giuffrida, M.C. / 2017 / Integration of isothermal amplification methods in microfluidic devices:Recent advances / Biosens. Bioelectron 90:174~186
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