Hello EvolDir community: A little while back I posted a message asking for suggestions for ethidium bromide alternatives.   Thank you very much to all who responded!   Below you can find a summary of responses, followed by my original post. It seems that most people still use ethidium bromide, though there are a few alternatives.   SYBR Safe (Invitrogen) and GelRed (Biotium) were by far the alternatives recommended most often, though there are of course some concerns associated with each.   Potential concerns include:   1) Stability.   SYBRSafe is light sensitive and degrades rapidly, so you need to add it fresh each time.   SYBR Green and SYBR Gold are more sensitive, but potentially more hazardous. GelRed can be used multiple times and seems to have similar stability to ethidium bromide.   2) Cost.   SYBRSafe is more expensive than other alternatives (though dilution may be an option).   the need to buy a different filter for SYBRSafe to capture optimal fluorescence (or make sure your new gel doc system comes with an appropriate filter).   Some have had success with the standard ethidium filter. GelRed uses the standard filter.   3) Sensitivity.   Some found a lack of sensitivity to detect bands <20 0bp with SYBRSafe (though others did not mention any problems).   4) Ease of use.   SYBRSafe and GelRed can be directly incorporated into the gel, whereas SYBRGold and SYBRGreen cannot (seems to affect mobility).   Also SYBRSafe should be added after the molten gel is allowed to cool for awhile. Other stains mentioned included SafeView (NBS Biologicals; noted as troublesome for cloning),   Envision (Amresco), GoldViewT ( http://www.sbsbio.com/eng_shiji.asp ), and Megafluor (Gentaur; noted for low sensitivity). Several people point out that anything that binds to DNA should be treated with caution, and some mention that the hazards of ethidium bromide may have been overstated (see http://rrresearch.blogspot.com/2006/10/heresy-about-ethidium-bromide.html and http://bitesizebio.com/2008/01/28/warning-dihydrogen-monoxide-is-worse-than-ethidium-bromide/ Recommendations for image capturing included Kodak, UltraLam, Eagle Eye, BioRad, BioVision 3000.   Also Kris Hundertmark's web page on gel docs comes highly regarded by myself and others: http://users.iab.uaf.edu/~kris_hundertmark/Lab/Geldoc.html .   Invitrogen's blue transilluminator was suggested a few times to avoid using UV altogether.   A design flaw in the system allows for buffer to seep in, but covering it with a sheet of acetate seems to solve the problem. Many people wanted to know about my super-awesome recipe for sodium-borate (SB) buffer.   It is from a BioTechniques paper a few years ago" Brody and Kern. 2004. BioTechniques 36: 214-216.   I follow the recipe as in the paper, making a 20X stock solution and diluting it down as needed.   An insider tip: it is helpful to use a fine-grained boric acid; otherwise it will take a long time to go into solution.   As a conductive media, it mitigates the runaway positive feedback loop between temperature and current that occurs with Tris-based buffers, thereby allowing super-fast separation of fragments (without the unfortunate side-effect of melting your gel).   When I experimentally tested TBE, TAE, and SB in the lab, I found similar resolution of both small and large products, and also calculated the cost of SB as about one-fifth that of TBE.   I have used it for routine separation and for cutting bands out of a gel.   It works fine with Qiagen's gel extraction kit" I haven't tried other kits. Original post: I have recently started a new lab and would like to explore less toxic alternatives to Ethidium Bromide staining of agarose gels. Although I am aware of SYBRSafe, I have not used it. Also, I have heard about dyes like Crystal VU (methyl violet stain from Genlantis) that do not require UV to visualize, but I think sensitivity suffers. Thus, I would like to poll the EvolDir community for their experiences and recommendations. Certainly, any chemical that binds to DNA should be handled with caution, but I am hoping to reduce toxicity and minimize accumulation of hazardous waste. Cost and sensitivity are of concern, although for the most part I am just 'checking' genomic DNA, PCR products, etc. Additionally, it would be convenient if the solution could be incorporated directly into the agarose gel (no post-staining). Also, I typically use SB (sodium borate) buffer for agarose gels (a switch from TBE or TAE that I highly recommend - lower cost, equally effective, and not exothermic like tris-based buffers so you can run gels faster - but that's a story for another day). Has anyone tried these stains with SB buffer? I also would be interested to know what gel documentation systems (homemade or commercial) people are using to visualize these gels. Any that come highly recommended (or that I should run away from)? I appreciate your candid comments and will of course compile and post responses. Thanks, Emily Emily K. Latch Assistant Professor Dept. of Biological Sciences University of Wisconsin - Milwaukee 3209 N. Maryland Ave. Milwaukee, WI 53211 Email: latch@uwm.edu Tel: 414-229-4245