I don't presume to know what your intentions are for seeking a non-toxic alternative specifically, but this blog link will help put some perspective on your query:

http://bitesizebio.com/articles/ethidium-bromide-a-reality-check/

The perfect is the enemy of the good in this case. I once had a post-doc who wouldn't even take the time to put on gloves to fish out his gel, which would have been much to the chagrin of our principle should he have ever discovered this deviation from protocol. All this is to say, people's opinion will vary -- in adhering to a minimum of PPE and common sense with EtBr, you should be fine using accepted processing methods in your laboratory.

SYBR® Green I Nucleic Acid Gel Stain

My lab has been using GelRed (Biotium) with good success. It's more sensitive that EtBr and since it's a 10,000x stock it's cheap enough, unless you're running a large number of gels continuously. In that case it's hard to beat EtBr which costs around $1 per milliliter at our institute.

We use Safeview in the lab (http://www.nbsbio.co.uk/product.asp?pID=2490&c=33551) It's quite good for regular PCR products, but not for larger (over 1kb) fragments.

We've used SYBR Gold and it works great.

Http://bitesizebio.com/articles/ethidium-bromide-the-alternatives/

GelRed DNA Stain (from Biotium) is better than SYBR Safe (from Invitrogen, now Life technologies). Also Gelred is stable in microwave ovens. We use iit directly on loading buffers.

Safeview, SYBRsafe, Gel Red / Gel Green and Megafluor are the safe alternatives of EtBr. I have used GelRed for my experiments.

I´ve tried GelRed (www.biotium.com) and worked fine!

I'd love there were a 100% secure DNA staining dye. Unfortunately, DNA binding capacity and non-toxicity are somewhat incompatible things. You can use SYBR which is less toxic, but should wear gloves and long sleeves exactly while using bromide. The greatest advantage is for the environment, certainly.

We use RedSafe and RealSafe, there are very similar and work fine.

We use GelRed, it is fine. Good quality staining, perhaps nicer than BrEt. But as Gustavo Santos said, "DNA binding capacity and non-toxicity are somewhat incompatible things". We protect ourselves from the dye (gloves and all that) but we don´t spend as much money in bromidium ethide diposal.

http://www.bio-rad.com/webroot/web/pdf/lse/literature/Bulletin_4110153A.pdf

We are using SYBR dyes and they work fine for most applications. But staining is always kind of toxic. Use personal protection and reduce the concentration of your dyes, sometimes more effective for staining and for costs.

Gel Green. It's very user friendly

you can Syber safe

You can use "gel green Nucleic acid stain" by Beckman coulter INc.

Gel green and Gel Red appear to be very costly. 500uL costs around 100$. It will be enough for only 500mL of Agarose gel. Am i correct?

SYBR gold nucleic acid gel stain from invitrogen is good

SYBR green is certainly an option but it is way more expensive and it needs quite a ot to get a view of a DNA band whereas EtBr you need way less concentrated.

I suggest GelRed. It's very user friendly but expensive and prone to deterioration. It's true: "DNA binding capacity and non-toxicity are somewhat incompatible things!"

I used GelRed (Biotium, Inc.) and it worked well for me. Another ethidium bromided alternative product is EZ Vision 6X Gel Dye (Amresco, Inc.). This product is just a gel dye that fluoresces under UV light as opposed to a stain. I tried it but did not like it as much as GelRed since it seemed to inhibit the DNA mobility. I have not used it for a long time so they might have improved it. You can call the company and ask for a free sample to try out before you buy it.

We have been using SYBRSafe (Invitrogen)...and it works very well!

SYBR safe and RED safe are non-toxic and both are working fine for most DNA and PCR products.

SYBR Safe DNA gel stain from Invitrogen cat# S33102

SYBR green is the best, but it is really expensive. Before make your choice, pay attention to the DNA mobility. I used to use the blue green loadying dye I and it changes the way DNA migrate (fragments looks longer than they are in the gel, and ofetn the bands show a "S" shape). It looks like it's a common problem among acid nucleic dyes..

Maybe you can use silver nitrate, is a classical method

SYBR gold works well, and considered to be less mutagen.

We used Gel Red for our huge TGGE gels (which have to be post-stained), but had mixed results with the quality of the staining. Sometimes it was really good, but other times we had a lot of large, ugly deposits on the gel (always the ones we wanted to publish!). Heating the stain does help reduce spotting, as suggested in the product literature, but we realised that one of our Gel Red tubes was simply a bad batch. When we complained, the folks at Biotium were less than helpful, never sent a replacement tube, basically tried to blame us for incompetence (!), and we lost a lot of time and money as a result. So, beware! We have now reluctantly gone back to Ethidium Bromide, and we use the "destaining tea bags" to decontaminate our solutions.

I use RedSafe (Ref. 21141, Intron Biotechnology). You will see green bands under UV light, but if you are running more than 100 ng DNA/well you can visualize the DNA as orange bands under visible light without the inconvenient of dimers formations due to UV light. It is really great.

I am using Syto9 (Invitrogen Life Technologies), and it works really good

We use SYBR safe at our lab. It works well with both UV light and with the blue light transilluminator from invitrogen (G6600) which reduces DNA damage if you need to use it for downstream applications.

I'll add another vote to GelRed by Biotium.

To answer your follow-up question on cost - yes, it's high. You'll use 1ul for every 10ml of gel. Remember, though, that you should realize some savings back by being able to reduce your reaction volume (due to increased staining sensitivity). If your end game is solely to show absence / presence of a product in your gel, you might go even cheaper and re-caste agarose gels a few times. The stain is heat tolerant and you need only to replenish it by about a third to a half of what would be normally called for. Use your new gels only for the publication quality photos.

I am using EZ vision dye, Its good for shorter runs, but the florouscence disappears on longer migration

I use safeview of ABM.

http://www.abmgood.com/SafeView-Plus%E2%84%A2-G468.html

It works very well.

My lab team work uses the SBR Green and we have had good results. The SBR Green is considered to be safe, less mutagen. Though the cost of this product is little high, I suggest you to try to use it (or test it) considering it to be a safer product.Good luck!

Gel-red is alternative

We have been using Gel-red for more then 3 years now and it works perfectly, at least as good as ethidium bromide, and considering all the advantages of being nom-toxic, i really do not think it is more expensive.

you may like non-toxic fast blast DNA stain or SYBR safe stain, but you consider first which sensitivity you need, and which visualisation equipment you have, software analysis

We have used SYBR safe from Invitrogen for the last 4-5 years. It works as well with UV light as ethidium. It can be used with blue light transilluminator and orange glasses/filters to reduce DNA damage if you need to gel purify etc. Really strong bands can even seen under visble light.

EMA, is cheap& safe

GelRed is pretty much a direct replacement, just as easy to use as EtBr. Phenix carries it, and it's not too expensive.

We testes quite a number of different dyes. Especially SYBR shows some change in the mobility of the DNA (so it seems to be of different size). HDgreen works best for us.

DEFINITELY GelRed (Biotium). You can use it directly on youy loading buffer (3 ul of GelRed per 1 ml of loading buffer with any tracking dye) and you will never have to buy it again. Use it for both your samples and ladder; you will see that the gels will have a superior contrast, without spending large amounts of staining reagent, and also it will be easy to reuse your gels again and again (even ten times!) Check it out and share your experience. And more important, you will not notice a big effect on DNA mobility (because ladder fragments are equally treated as your samples)

SYBR safe DNA gel stain from Invitrogen

you may use Gelred

Hello everybody!

I am following some topics, now just for fun. More precisely, it's a tragi-comic situation, isn't it.

Let's be honest, and let's admitt it, don't you think some of us post an "answer" just to post something here. Or just attempting to get higher scores on this gate, perhaps.

Dont you guys have something more productive to do? GelRed - 22 mentions in 56 answers, until Veera noticed it might be costly. SYBR dyes - 21 mentions, safe 8, gold 3, make sure Verra got the message. Appreciated whether some arguments follow (such as in Raul's note, Louise's and others)

Although all of us are concerned about our safety as well as protection of the environment, such an efervescence...still...

I don't presume to know what your intentions are for seeking a non-toxic alternative specifically, but this blog link will help put some perspective on your query:

http://bitesizebio.com/articles/ethidium-bromide-a-reality-check/

The perfect is the enemy of the good in this case. I once had a post-doc who wouldn't even take the time to put on gloves to fish out his gel, which would have been much to the chagrin of our principle should he have ever discovered this deviation from protocol. All this is to say, people's opinion will vary -- in adhering to a minimum of PPE and common sense with EtBr, you should be fine using accepted processing methods in your laboratory.

You may consider DAPI or propidium iodide. Both are non-toxic.

You may use 1x Syto9 (Invitrogen)

We just did a comparison of different methods looking for the best signal. We used GelRed, RealSafe, SyberSAfe and EZ-Vision, all non-toxic methods.

After 45 minutes running and 3 UV exposition, GelRed and RealSafe still showed very good staining. The bad thing is the price, both of them cost double than SyberSafe for instance.

I hope this is useful for you.

Gel green from phenix works very well for us

GelStar from Lonza is also very good

http://bio.lonza.com/uploads/tx_mwaxmarketingmaterial/Lonza_ManualsProductInstructions_GelStar_Nucleic_Acid_Gel_Stain_-_Protocol.pdf

Yes, SYBR safe DNA gel stain from Invitrogen is a a non toxic DNA staining dye.

Sybr safe has been suggested and I would second this. However, use more then the reconmended volume for optimum staining. We typically use 5ul for 30ml gels and 15ml for larger gels.

This is assuming you are refering to agarose gels. If you are staining other types of gels, sybr gold is great. It stains single stranded to so capture the integrity of your RNA!

I use gel red from Invitrogen for visualisation of dna bands. It's simple and safe.

Midori Green Nucleic Acid Staining Solution from Bulldog:

www.bulldog-bio.com/midori_green.html

I have used Gel Green, and now I use Midori Advance Green. Is cheaper, but gives good results.

It might be worth pointing out that depending on what you are doing you may not even need to stain your DNA to see it in the gel. DNA itself absorbs UV light strongly... there are many tricks based on this which allow you to see your bands unstained. However, 1) UV exposure will of course do some amount of damage to the DNA and 2) UV shadowing is much less sensitive than staining.

I use 0.2% methlyene blue in H2O for any DNA I want to extract from the gel (very cheap). It is not a sensitive as EtBr and has a higher background, but since you use fluorescent light to visualize the DNA (it does not intercalate in the DNA so no fluorescence like EtBr) there is no breaking of the DNA as there is with EtBr and UV "at any wavelength". Since it does not intercalate in DNA it is not mutagenic (it binds to the DNA backbone).

I also use methlyene blue in all my water baths to prevent microbial growth, add just enough to turn the water medium dark blue (it takes a few minutes for it to dissolve completely so add small amounts slowly with stirring).

We've been using SYBR safe with success, but you may need to optimize the volume to obtain good staining.

Good luck!

There are some dyes which are commercially available for staining gel... called GelRed.

http://www.phenixresearch.com/Images/RGB4102_GelRed_DMSOProductSheet.pdf..

We use GelRed from Biotinuml and it works good; we used EtBr previously, then we switched to GelRed. For 120 ml of 2% agarose gel we use 12,5 μl of GelRed.

Any phenothiazine flourochrome will do since it will intercalate between the bases at high concentration with a sufficiently high binding constant to remain in situ during handling. The use of DNAse will of course eliminate the association and hence a built in control can be established if needed. Suggest the following papers:

1. Crémieux A, Chevalier J, Sharples D, Berny H, Galy AM, Brouant P, Galy JP,

Barbe J. Antimicrobial activity of 9-oxo and 9-thio acridines: correlation with

interacalation into DNA and effects on macromolecular biosynthesis. Res

Microbiol. 1995 Jan;146(1):73-83. PubMed PMID: 7538688.

2: Berny H, Bsiri N, Charbit JJ, Galy AM, Soyfer JC, Galy JP, Barbe J, Sharples

D, Mesa Valle CM, Mascaro C, et al. 1,4-Dimethoxy-9(10H)-acridinone derivatives.

Synthesis, DNA binding studies and trypanocidal activity. Arzneimittelforschung.

1992 May;42(5):674-9. PubMed PMID: 1530683.

3: Sharples D, Spengler G, Molnár J, Antal Z, Molnár A, Kiss JT, Szabó JA,

Hilgeroth A, Gallo S, Mahamoud A, Barbe J. The interaction between resistance

modifiers such as pyrido[3,2-g]quinoline, aza-oxafluorene and pregnane

derivatives with DNA, plasmid DNA and tRNA. Eur J Med Chem. 2005

Feb;40(2):195-202. Erratum in: Eur J Med Chem. 2005 Oct;40(10):1070. PubMed PMID:15694654.

4. Wagenknecht HA. Fluorescent DNA base modifications and substitutes: multiple

fluorophore labeling and the DETEQ concept. Ann N Y Acad Sci. 2008;1130:122-30.

Epub 2007 Dec 20. Review. PubMed PMID: 18096856.

5: Latt SA. Fluorescent probes of chromosome structure and replication. Can J

Genet Cytol. 1977 Dec;19(4):603-23. Review. PubMed PMID: 76502.

Also, the acridines are relatively non-toxic and inexpensive, and their disposal is not a problem

Recently, people in my lab have used Green-Glo - and they're pretty happy with it!

Am also used GelRed since 2010 with good results that you could find on our articles on the Implementation of National reference Lab of Buruli ulcer in Togo.

Midori Green DNA dye from Nippon Genetics is hands down one of the best alternatives to EtBR. It has high sensitivity and its not that expensive. I use it myself and am very much satisfied with it.

Cheers,

Sudip

I've been using GelRed quite happily as well.

Just an update on my former comment. We just change from SybrSafe to RealSafe. It costs double-prize but we checked the staining protocol using 50% of the suggested volume and it still labels better than SybrSafe.

same with the later answer, Gelred by Biotium is the best Etbr replace and i always use gelred in my research now.... and gelred is better than sybr and other

Hi in our laboratory we changed from Etidium to SybrSafe, just changing the filter. In this way we saved time (around 1/2 hour) and the risky manipulation of the etidium

SybrGold does a very good job and has better sensitivity (at least 25x), and works on ss and ds nucleic acids

GelRed is pretty good but too expensive, this is what we are currently using when our virolab turned Ethidium free. You may use pre or post staining protocol and you are safe at least from what is currently known.

In our lab we use SYBRSafe, which works fine enough for our standard molecular biology. I'm guessing that RNA-research might need higher sensitivity.

But... how anyone can claim that a molecule binding / intercalating DNA is not mutagenic, is beyond my understanding. So I don't know what a "non-toxic DNA staining dye" would actually be! I.e. use your gloves. ;-)

In our lab we use GELRED and it´s preety good

Andrea,

The dyes that intercalate are mutagenic because in the presences of UV they break the DNA backbone (and likely even in the absence of UV, but at a lower rate). However, there are dyes like methylene blue that predominantly do not intercalate in between the DNA bases, but instead bind in the DNA grooves, and this allows it to be seen with visible light.

SyBr Green is a very good, commonly used alternative to ethidium bromide

Gary,

Thanks for explaining the mechanism, I hadn't thought about it. So you would say that these dyes would not interfere with DNA function? I just have a bad feeling about anything binding DNA, although I do understand that there is a big difference between an intercalating substance and one binding the surface.

You can try Cyber Safe of Life technologies and Safe blue of Bangalore Genei. you can check the following weblinks for more details.

http://www.merckindiawebapps.com/merckindiawebsites/merckmillipore/index.html#/221/zoomed

http://www.invitrogen.com/site/us/en/home/Products-and-Services/Applications/DNA-RNA-Purification-Analysis/Nucleic-Acid-Gel-Electrophoresis/DNA-Stains/SYBR-Safe.html

You can prove gel-red.

Gelred or gelgreen.

We use GelGreen and are very satisfied. I believe it excites better with blue light than UV, but we still image on a UV box and have good results. If you do get a blue light box, cutting bands out is amazing. You can see the DNA very easily, and there is no rush to get the cutting done.

I remember to have used oneof the Hoechst compounds for live-imaging. Look in Life Tech web site for it.

SYBR green is nowadays preferred

I recently used Midori green DNA stain. It is non toxic and is as sensitive as Ethidium bromide and can be used exactly the same way in agarose gel electrophoresis. You can refer the given link

http://www.nippongenetics.eu/dnarna-electrophoresis/dna-stains/midori-green-dna-stain/

SyBr Green is less toxic and good alternate to Ethidium bromide

Gel imaging and nucleic acid binding dyes are widely used in today’s life science

laboratories to visualize DNA fragments in agarose gels. Ethidium bromide (EtBr)

has been the predominant dye used for nucleic acid gel staining for decades

because of its low initial price and generally sufficient sensitivity. However,

the safety hazard and costs associated with decontamination and waste disposal

can ultimately make the dye expensive and unsafe to use. For this reason, safer

alternative gel stains were developed by scientists at Biotium, Inc. GelRed™ and

GelGreen™ dyes are a new generation of fluorescent nucleic acid gel stains

designed to replace the highly toxic EtBr. Three attributes make GelRed and

GelGreen dyes superior to EtBr and other EtBr alternatives: low toxicity, high

sensitivity, and exceptional stability.