Gelred, Gelgreen, SYBR Safe and Ethidium Bromide (Etbr) in Nucleic Acid Gel Staining

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NT-BQ041U Overview of DNA Gel stains: GelRed, GelGreen, SYBR Safe and Ethidium Bromide (EtBr) in Nucleic Acid Gel staining GelRed GelGreen SYBR Safe EtBr Note Sensitivity +++++ +++++ ++ +++ See comparison of GelGreen vs. SYBR Safe done by Clare Chemical Research who makes the visible light transilluminator http://www.clarechemical.com/gelgreen.htm Photostability +++++ +++++ +++ ++++ Mutagenicity Non-mutagenic Non-mutagenic Non-mutagenic Highly mutagenic! (Ames Test) (diluted) Speed to enter cells Does not enter cells Does not enter cells Enters into cells Can enter cells rapidly Cytotoxicity No No Yes Yes Waste disposal in drain Not tested Not tested Yes No! Needs to be collected for special disposal Compatibility with UV Yes Yes Yes Yes GelRed and EtBr are spectrally very similar. excitation Compatibility with 488nm Not optimal Yes Yes Not optimal Ar laser excitation or Dark Reader Compatibility with EtBr Yes No. A green filter is No. A green filter is Yes GelRed and EtBr are spectrally very similar. filter needed needed Detection of ds-DNA, ss- Yes Yes Yes Yes DNA, RNA in agarose gel Detection of ds-DNA, ss- Yes No Yes Yes DNA, RNA in polyacrylamide gel Compatibility with heating Yes. Can be done Yes maybe, only briefly Yes. Can be done GelRed is nearly as stable as EtBr by microwaving repeatedly repeatedly Dye removal by ethanol Yes Yes Yes Yes precipitation Compatibility with cloning Yes Yes Yes Yes Need for adding dye to No No Yes Yes GelRed and GelGreen do not run significantly in the running buffer for direction opposite migrating DNA while SYBR Safe maximal sensitivity and EtBr both do. Need for protection from Not necessary Recommended Recommended Not necessary light during gel staining Storage temperature Room temperature Room temperature Room temperature Room temperature Cost Economical Economical Economic (1X) to Inexpensive to buy but GelRed and GelGreen offer the best inexpensive expensive to dispose performance/price ratio (10000X) For any information, please ask : FluoProbes® / Interchim; Hotline : +33(0)4 70 03 73 06 GelRed™ and GelGreen™ are trademarks from Biotium. SYBR Green™ is trademark of Invitrogen Rev.I03E [email protected] P.1 Contact your local distributor.

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Agarose Gel Electrophoresis
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IMagING Molecular Imager ® Gel Doc™ XR+ and ChemiDoc™ XRS+ Systems ■■ Reproducible results Imaging Fluorescently Stained DNA with Alternatives to ™ ■■ Safe■ DNA imaging Ethidium Bromide Using the XcitaBlue Conversion Screen ■■ Flexible and easy to use platform ■■ Extra resolution Introduction EtBr Ex. EtBr Em. SYBR ® Safe Ex. SYBR ® Safe Em. and quality images Ethidium bromide (EtBr) is the most commonly ■■ Trusted name used fluorophore for staining DNA due to its availability and low cost. However, it is a powerful mutagen and requires special waste disposal procedures. Furthermore, EtBr is optimally excited by UV light (Figure 1), which is known to damage DNA via thymine dimer formation and strand breaks. This in turn leads to decreased efficiency in cloning and transformation (Paabo et al. 1990, 250 300 350 400 450 500 550 600 650 700 750 800 Grundemann and Schomig 1996). Therefore, it is desirable to develop the use of nonnoxious and GelGreen Ex. GelGreen Em. environmentally friendly technologies. The recently developed GelGreen (Biotium, Inc.) and SYBR® Safe (Invitrogen Corp.) stains exhibit low mutagenicity and no toxicity as reported by their manufacturers (refer to safety information on manufacturer’s Web sites). These stains absorb optimally in the blue region of the spectrum (410–510 nm), emit in the green region (see Figure 250 300 350 400 450 500 550 600 650 700 1), and do not require DNA-damaging UV Wavelength, nm excitation. They have sensitivity equal to or greater Fig. 1. Excitation (Ex.) and emission (Em.) spectra of than that of EtBr, although they may require ethidium bromide, and SYBR ® Safe stain (top panel) slightly longer exposure times.
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