NEW StainIN™ eco-RED Nucleic Acid Stain - a less toxic alternative to ethidium bromide
Applications
- Staining of DNA or RNA in agarose and polyacrylamide gels
- ssDNA, dsDNA and RNA visualization under the UV light
- ssDNA, dsDNA and RNA visualization under LED light
Benefits
- Replaces discontinued StainIN™ RED Nucleic Acid Stain NAS0101
- Safe, DMSO-free, economical alternative to ethidium bromide
- Universal – in gel staining, or post- run staining, no de-staining.
- Ambient temperature storage (protected from light)
StainIN™ eco-RED Nucleic Acid Stain
Product information "StainIN™ eco-RED Nucleic Acid Stain "
The carcinogenicity and a high toxicity of a widely used gel stain Ethidium Bromide (EtBr) is one of the biggest safety and environmental problems in the labs where gel electrophoresis is a daily routine. Therefore, there is a need for a safer ethidium bromide alternatives to be used in laboratories. The novel less toxic nucleic acid dyes used for agarose gel electrophoresis address this ethidium bromide problem and minimize environmental concerns.
highQu StainIN™ eco-RED Nucleic Acid Stain provides a significantly safer alternative to ethidium bromide while delivering up to several times higher detection sensitivity. Same like ethidium bromide, the stain is used during the process of electrophoresis in an agarose gel or PAAG and (optionally) in the electrophoresis running buffer, or alternatively, for a post-run staining of the gels in a tray with the buffer.
NEW: Download the Nucleic Acid Gel Staining Troubleshooting Guide
Despite having a proven lower mutagenicity, all nucleic acid stains shall be handled with care, as they interact with DNA and RNA. Used electrophoresis buffer with a stain as well as melted gels shall be run through approved (carbon) filters. If the absence of residual fluorescence is confirmed, the liquids can be disposed with plenty of water down the drain.
Alternatively, all used liquids can be autoclaved or heated as the StainIN™ eco-RED Nucleic Acid Stain degrades during the long exposure to above 100ºC temperature.
The safety office shall be consulted periodically to match local regulations, as these regulations vary and change. Despite of tests described in this report, the seller of the listed stains and other reagents does not take any responsibility for the possible damage resulting from handling any sold chemical or reagent.
Frequently asked questions about the new Nucleic Acids Stain
1. What are the advantages of the new StainIN™ eco-RED NA Stain over the old version StainIN™ RED?
- The new stain is purely aqueous solution. The old StainIN™ RED had DMSO as solvent.
- The new stain is room temperature stable, shipped at ambient temperature. Store it at your electrophoresis bench. The old StainIN™ RED was stored at +4°C.
- StainIN™ eco-RED NA Stain is more flexible – detectable under both UV and LED light.
- More convenient protocol – the stain can be used both during the gel run and as a post-run staining.
- Same or better sensitivity compared to the old stain.
2. Is the color of the solution the same as before?
The new stain is a darker shade of red.
3. What is the concentration of the StainIN™ eco-RED Nucleic Acids Stain?
It is 10,000X concentrated, however the number of applications for post run staining is difficult to calculate because multiple gels can be stained in the same staining solution.
4. What is the solvent used in the StainIN™ eco-RED Nucleic Acids Stain?
The solvent is 100% water.
5. Which stains are similar to StainIN™ eco-RED Nucleic Acids Stain?
The closest similar stains are Ethidium Bromide and GelRed (Biotium).
6. What is the sensitivity of the StainIN™ eco-RED Nucleic Acids Stain in ng/band?
The sensitivity ranges from 0.1 to 0.3 ng DNA/band or even higher, depending on factors such as the staining method used, imaging capability, detection, and gel thickness.
7. What is the emission wavelength of the StainIN™ eco-RED Nucleic Acids Stain?
The emission wavelength is 600 nm.
8. What is the excitation wavelength of the StainIN™ eco-RED Nucleic Acids Stain?
The excitation wavelength is 300 nm.
9. Can StainIN™ eco-RED Nucleic Acids Stain be used for in-gel and in-buffer staining?
Yes, it can be used for both in-gel and in-buffer staining post electrophoresis run.
10. How should the stain be used during a gel run in agarose gels?
Use 8-10 µl of the stain in 100 ml agarose gel solution cooled down for pouring, nothing to be added in to the electrophoresis buffer.
11. How to use StainIN™ eco-RED Nucleic Acids Stain with PAAGE gels? Same like with agarose gel, or using the post-run staining protocol.
12. Can the StainIN™ eco-RED Nucleic Acids Stain be detected under UV light?
Yes, it can be detected under UV light, with ethidium bromide or GeRed filters.
13. What color does the stained DNA appear as under UV light?
DNA appears as red-orange under the UV light.
14. What color does RNA appear as under UV light?
RNA appears as red-orange under UV light.
15. Does StainIN™ eco-RED Nucleic Acids Stain color both single-stranded and double-stranded DNA?
Yes, it can stain both single-stranded and double-stranded DNA, and RNA.
16. Can the stained DNA be detected with a blue LED?
Yes, the StainIN™ eco-RED Nucleic Acids Stain -stained DNA can be detected with a blue LED.
17. Is StainIN™ eco-RED Nucleic Acids Stain compatible with cloning?
Yes, as it can be used with LED without damaging DNA.
18. Has StainIN™ eco-RED Nucleic Acids Stain undergone toxicity testing? Yes, it is less toxic than Ethidium Bromide See the toxicity testing report.
19. Can StainIN™ eco-RED Nucleic Acids Stain be used for post-run staining?
Yes, it can be used for post-run staining.
20. Can the staining buffer (electrophoresis buffer with the NA stain) be reused?
Yes, the staining buffer can be reused several times, depending on gel thickness and the amount of DNA stained. It should be stored in the dark in a glass or plastic container. To avoid StainIN™ eco-RED Nucleic Acids Stain absorbance on plastic or glass over the prolonged storage of more than 24 hours, make a fresh staining solution every day.
21. Is the StainIN™ eco-RED Nucleic Acids Stain sensitive to light?
Yes, same as all other fluorescent nucleic acid’s stains, the StainIN™ eco-RED Nucleic Acids Stain is light-sensitive and should be protected from light.
22. Can StainIN™ eco-RED Nucleic Acids Stain be stored at +4°C?
The recommended storage conditions for this stain is ambient temperature at about +20°C.
However, due to common cooled shipments, it can be exposed for a short time to +4°C. If accidentally shipped with cooling packs at +4°C, simply mix the product to ensure the stain is completely dissolved. And keep it further only at ambient temperature.
23. What happens if the StainIN™ eco-RED Nucleic Acids Stain was put or exposed to -20°C and was frozen?
If frozen, the StainIN™ eco-RED Nucleic Acids Stain should be disposed and the new product shall be ordered. Do not use it if once frozen.
- Download Protocol and Specifications - Product Insert StainIN™ eco-RED Nucleic Acid Stain
- NEW: Download the Nucleic Acid Gel Staining Troubleshooting Guide
- Ask for a Sample today
- Have technical questions? Contact us
- Download a list of Publications mentioning highQu products
- Or see how others use our products at bioz.com or google scholar
Nucleic Acid Stain Troubleshooting Guide For most common agarose gel staining issues
NOTE:
ü In case of any problems appearing with the gel quality, please read the product manual carefully again to refresh the recommended protocols and important product usage tips.
ü Compare the recommended protocols with the ones you use and draw appropriate conclusions. Repeat the experiments with fresh buffers/gels by following protocols strictly as recommended.
The most important product properties and protocols to follow:
Storage:
ü Store the stain strictly at +15–25 °C in the dark.
ü The stain is sensitive to light and should be protected from light all the time during storage, usage and gel staining. Keep the trays with the staining buffer protected from light f.e. covered during the staining and between stainings.
ü The stain is stored at ambient temperature at about +20°C. Due to common cooled shipments, it can be exposed for a short time to +4°C. If accidentally shipped with cooling packs at +4°C, warm the stain solution shortly to maximal 30°C and mix well to dissolve all precipitate. And keep it further only at ambient temperature.
ü Never freeze the product. Discard if frozen. If once frozen, the new product shall be ordered.
Usage:
• Use 8-10 µl of the stain solution per 100 ml of the gel right before casting the gel.
• Add stain to the gel solution only after agarose cools to hand-warm temperature (~40–50 °C) and mix well.
• Optionally, if low NA concentration is expected, add 2-5 µl of the stain solution per 100 ml of the 1X electrophoresis running buffer.
• Gel destaining is not needed, but it might help to reduce the background. For optional destaining, keep the stained gel in fresh water or 1X electrophoresis buffer batch while slowly shaking for 10 minutes.
• If you reuse the molten gels, add at least half a portion of the stain each time after boiling and cooling the gel solution down. Reused gels might have higher background staining.
• For post-run staining, soak the gel for 10-30 minutes into the 100 ml solution of 1X electrophoresis buffer freshly mixed with 10 -30 µl of the stain. The amount of the stain can be adjusted, as the staining intensity depends on gel percentage and thickness. Thick and high percentage gels require more stain and longer staining time.
• Keep the tray protected from light (f.e., covered) during the staining and between stainings.
• The same staining solution can be used for up to 5-10 gels, or up to the sensitivity is acceptable.
• Longer staining time of 30 minutes gives better results when detecting low DNA amounts, however, it may cause background or diffusion of smallest NA fragments.
• To avoid stain absorbance on plastic or glass during the storage, make a fresh staining solution every day.
Short Nucleic Acid Stain Troubleshooting:
|
Problem |
Likely Cause |
Solution |
|
|
DNA bands too faint, low staining sensitivity |
1. Stain degraded due to cooling/freezing or light exposure 2. Stain not mixed well before adding 3. Stain added to too hot agarose and inactivated 4. Insufficient stain amount or too short post-run gel staining time 5. In case of post-run staining, stain absorbed by a bad quality plastic of the staining tray 6. Weak or uneven stain distribution in the agarose or the stain was partially precipitated 7. No dye added to running buffer when DNA concentration is low |
1. Store at +15–25 °C in the dark; discard if frozen. Keep trays with staining buffer protected from light during the staining and between stainings 2. Mix gently and thoroughly before every pipetting 3. Add stain only after agarose cools to hand-warm (~40–50 °C) 4. Increase stain volume slightly (if you used minimal amount) or extend post-run staining for a few min 5. If possible, use high quality non-binding plastic trays for staining the gels. If the staining buffers shall be reused several times, protect from light 6. Warm stain shortly to 30°C if precipitate is visible and mix until fully homogeneous. Mix the stain well in agarose solution or staining buffer by slight shaking. 7. Add the stain not only into the gel solution, but also use 2–5 µl per 100 ml electrophoresis running buffer for low-abundance samples |
|
|
Smeared or distorted bands |
1. Stain added to very hot agarose (damaged) 2. Uneven stain distribution in precast gel 3. DNA overloaded or contaminated with proteins or nucleases 4. Electrophoresis voltage too high 5. Gel-wells were not formed well or were leaky |
1. Add stain only after agarose cools to hand-warm (~40–50 °C) 2. Mix gel solution gently and avoid bubbles 3. Reduce DNA amount loaded, increase the sample volume by adding sample loading buffer. If nucleases are suspected, change the sample loading buffer, and make new gel and new electrophoresis buffer. 4. Run the gel at ~110–130 V 5. Cast a new gel carefully and repeat the experiment 6. If problems persist, try rather post-staining instead of in-gel staining |
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|
Excessively high gel background |
1. Too much stain in gel or buffer 2. Stain exposed to cold or light 3. Thick gel or long staining time increasing background fluorescence |
1. Reduce stain amount by 20–50% for thin and low percentage gels 2. Keep gel and stain solutions protected from light at room temperature 3. Shorten post-run staining time to 10–20 min. Use optional destaining (soak in water or fresh electrophoresis buffer for 5–15 min) to reduce background |
|
|
Precipitation in stain stock |
1. Temporary exposure to cold (e.g., 4 °C) for example if the stain is used/stored in cold laboratory rooms |
1. Warm to +25–30 °C until dissolved; mix gently. If completely frozen at any point - discard the stain and order the new one. |
|
|
Weak fluorescence with reused agarose or reused buffer |
1. Not enough stain added after reheating the gel in case of gel staining 2. Stain degraded by repeated heating cycles in case of reusing the agarose 3. Stain used-up in case of reusing the same buffer with the post-run protocol for multiple gels stainings |
1. Preferably, always use freshly made gel with freshly added stain or fresh buffer for post-run staining. If reuse is wanted - add at least half the original stain dose after each re-melt of agarose. 2. Add stain only after agarose cools to hand-warm (~40–50 °C). But note- remelting and adding the stain will increase the background staining. 3. You can use the same staining buffer for several gels until the staining efficiency is enough. As the stain is binding to the DNA and gel, after several uses, add a little bit of stain again and use further. But note – the buffer shall be kept away from light exposure and only high-quality plastic or glass trays shall be used to avoid stain absorption. |
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- Download Protocol and Specifications - Product Insert StainIN™ eco-RED Nucleic Acid Stain
- Download MSDS StainIN™ eco-RED Nucleic Acid Stain
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- Download Protocol and Specifications - Product Insert StainIN™ eco-RED Nucleic Acid Stain
- Download MSDS StainIN™ eco-RED Nucleic Acid Stain
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- Or see how others use our products at bioz.com or google scholar
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