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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.

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Online
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1437-4331
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Volume 43, Issue 8 (Aug 2005)

Issues

Comparative analysis of the DNA staining efficiencies of different fluorescent dyes in preparative agarose gel electrophoresis

Qing Huang
  • Department of Laboratory Medicine, Southwest Hospital, the Third Military Medical University, Chongqing, China
/ Wei-Ling Fu
  • Department of Laboratory Medicine, Southwest Hospital, the Third Military Medical University, Chongqing, China
Published Online: 2011-09-21 | DOI: https://doi.org/10.1515/CCLM.2005.141

Abstract

Ethidium bromide (EB) is a mutagen and toxin that is widely used in the laboratory for visualization of nucleic acids. Safer nucleic acid stains, such as SYBR ® Gold, SYBR ® Green, GoldView™, GeneFinder ®, and GoldStar ®, have been developed. However, there has been no systematic comparative analysis of the staining efficiencies of these dyes. In the present study, SYBR ® Gold, SYBR ® Green I, GoldView™ and EB were compared. Although both SYBR ® Gold and SYBR ® Green alter electrophoretic mobility and thus DNA size estimates, they are cost-effective alternatives to EB. SYBR ® Gold was more sensitive than SYBR ® Green I at detecting short fragments, but 50-bp bands were clearly visible using either dye when visualized with a long integration time. SYBR ® Gold or SYBR ® Green I are sensitive and relatively safe alternatives to EB. In our laboratory, the SYBR ® Gold method is now used routinely by all members of our group with great consistency and success.

Keywords: agarose gel electrophoresis; ethidium bromide; fluorescent dyes; GoldView™; methods; SYBR ® Gold; SYBR ® Green

References

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    Lunn G. Decontamination of ethidium bromide spills. Trends Genet 1990; 6: 31.

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    Singer VL, Lawlor TE, Yue S. Comparison of SYBR Green I nucleic acid gel stain mutagenicity and ethidium bromide mutagenicity in the Salmonella/mammalian micro-some reverse mutation assay (Ames test). Mutat Res 1999; 439: 37–47.

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    Tuma RS, Beaudet MP, Jin X, Jones LJ, Cheung CY, Yue S, et al. Characterization of SYBR Gold nucleic acid gel stain: a dye optimized for use with 300-nm ultraviolet transilluminators. Anal Biochem 1999; 268: 278–88.

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    Miller SE, Taillon-Miller P, Kwok PY. Cost-effective staining of DNA with SYBR green in preparative agarose gel electrophoresis. Biotechniques 1999; 27: 34–6.

About the article

Corresponding author: Prof. Wei-Ling Fu, PhD, MD, Department of Laboratory Medicine, Southwestern Hospital, the Third Military Medical University, Chongqing 400038, China Phone: +86-23-6546-0802, Fax: +86-23-6546-0909,


Received: 2005-04-04

Accepted: 2005-06-17

Published Online: 2011-09-21

Published in Print: 2005-08-01


Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2005.141. Export Citation

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