SafeView™ Classic
Cat. No. | G108 |
Name | SafeView™ Classic |
Unit | 1.0 ml |
Category | SafeView™ DNA Stains |
Description |
abm’s SafeView™ Classic is a safe, improved formulation of nucleic acid stain for agarose gel electrophoresis. SafeView™ Classic directly replaces toxic Ethidium Bromide and eliminates its associated contamination risks with glassware, gel apparatus and environment. Features: |
LED Viewer Compatibility | Yes |
Application |
Safe Detection of dsDNA, ssDNA and RNA in agarose gels. |
Concentration | 10,000X |
Format General | Pre-cast or Post-stain |
Storage Condition |
Store at 18-25°C |
Note |
All SafeView DNA Stains are ISO-13485 certified. |
Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. G108 |
Can SafeView Classic be a replacement for ethidium bromide? Can I do gel extraction with it? | |
SafeView Classic can be used as a complete replacement for your ethidium bromide workflow. Yes, SafeView Classic can be used during gel extraction - in house testing demonstrated that gel extraction of DNA fragments using a SafeView Classic gel resulted in no decrease in ligation efficiency vs ethidium bromide. |
How does SafeView work and why is it not carcinogenic? | |
abm's SafeView products contain fluorescent compounds that have a strong affinity to DNA and RNA nucleic acids. Once bound to nucleic acids, the compound fluoresces under specific wavelength of light which can then be visualized using a standard UV/Blue light imager. There may be some unknown effects of SafeView products that have not been documented in literature but these would also apply to popular SYBRSafe as well; however, SafeView products are not as carcinogenic as ethidium bromide. |
How do I use SafeView products? | |
Safe-Red and Safe-Green are in a 6X loading dye format: mix samples and DNA marker with Safe-Red or Safe-Green at a 1:5 (dye : sample) dilution ratio, and load onto an unstained gel. SafeView Classic and Safe-Red Gel are in a 10,000X format: for pre-cast, add 10µl SafeView Classic or Safe-Red Gel per 100 ml molten agarose, mix gently and cast the gel; for post-stain, prepare and run an unstained agarose gel, next submerge the gel in post-staining solution of 30 μl SafeView Classic or Safe-Red Gel per 100 ml 1X TAE or 1X TBE buffer for 30min in the dark with light agitation, image accordingly. |
Can the SafeView products differentiate double stranded vs single stranded nucleic acids? | |
No, our SafeView products will bind to both double-stranded and single-stranded nucleic acids, albeit with lesser efficiency for single-stranded nucleic acid species.
|
At what temperature do I store the SafeView products? | |
SafeView Classic, Safe-Red and Safe-Red Gel should be stored at 18-25°C. Safe-Green should be stored at 4°C. |
Do I need a special filter for photography of DNA gels stained with SafeView products? | |
All SafeView products are compatible with UV or Blue Light imagers. However, Safe-Green and SafeView Classic are more optimally visualized under Blue Light, making these stains safer for the user and their samples.
|
How long does SafeView Classic stain last in a gel? | |
Our in-house testing has shown that SafeView Classic stained gels (>10ng DNA loaded per lane) can still be effectively visualized up to 1 week later with only a slight decrease in brightness. Gels should be stored properly to maintain a good signal, sealed in a plastic bag with wet paper towel loosely wrapped around the gel, away from light. Similarly, a pre-cast unused SafeView Classic gel can be stored and used up to 1 week after being prepared provided they are stored in ideal conditions (sealed in plastic bag, away from light).
|
What is the sensitivity of the SafeView products? | |
Safe-Green: 0.2-0.6 ng DNA per band Safe-Red: 0.6-1.0 ng DNA per band Safe-Red Gel: 0.6-1.0 ng DNA per band SafeView Classic: 1.0-2.0 ng DNA per band |
Can SafeView products be used to post-stain gels? | |
Yes, SafeView Classic and Safe-Red Gel can both be used for either pre-cast or post-stain gels.
|
Why is the EtBr signal stronger in the pictures when I compare SafeView with EtBr? | |
Most gel imaging systems have been optimized for EtBr so this is a likely reason why the EtBr signal may be stronger in the pictures.
|
Is there any guidance on how long the gel lasts once the stain has been added? | |
A pre-cast unused SafeView Classic gel can be stored and used up to 1 week after being prepared provided they are stored in ideal conditions (sealed in plastic bag, away from light). If it is stored longer than a week, there will be decreased resolution.
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How do I dispose Agarose Gels stained with Safeview and how do I dispose buffers and gloves? | |
Dispose SafeView™ Classic as you would any other non-carcinogenic fluorescent dye (eg. Acridine orange; Propidium iodide).
All gels and contaminated “non-sharp” lab debris (e.g., gloves, pads, towels, tubes, etc.) that are processed using this dye can be discarded in the trash.
Spent running buffer solutions and destaining solutions that contain the dyes can either be collected and disposed of through the HWMU or collected and run through an approved filter device.
The buffer solutions that have been run through the approved filter should be checked under the appropriate light source for complete removal of the dyes, and if it passes (does not fluoresce), the liquid can be disposed of down the drain with a copious amount of water as long as no other materials are present that would cause the material to be a Hazardous Waste.
The filters that have been used up and are no longer effective must be disposed of through the HWMU.
|
Will I need an additional loading buffer for my samples? | |
If using Safe-Green or Safe-Red, you will not need to add any additional loading dyes or buffers. If using SafeView Classic or Safe-Red Gel, you will need to add an appropriate DNA loading dye to your samples in order for samples to properly settle and stay in the wells during electrophoresis. |
Do SafeView products give problems in the process of cloning? | |
We recommend using SafeView Classic or Safe-Red Gel which can both be used for the gel extraction of DNA fragments destined for cloning. We have observed no decrease in cloning efficiency using these stains.
|
We see migrations and band shifting of our fragments when using Safe-Green. Are there any recommendations that you can give us to minimize this band shifting? | |
Band shifting to a certain degree is unavoidable for any nucleic acid stain as most of these fluorescent compounds are large, positively charged molecules. Loading dye format nucleic acid stains, while convenient, may also suffer from more prominent band shifting due to the stain binding and migrating with the sample simultaneously during electrophoresis. Post-staining, while takes longer to complete, may be the optimal choice of nucleic acid stain strategy if band shifting issue is a priority. |
I cannot see 100bp and 200bp bands on a 1% gel. What should I do? | |
It is often difficult to detect smaller 100bp and 200bp bands on a 1% gel especially if samples are of low concentration. We recommend visualizing smaller fragments on a higher concentration gel - ideally 2% agarose.
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Can I use SafeView products in polyacrylamide gels? | |
SafeView products are designed for use with agarose gels, and are not optimized for polyacrylamide gels. However, SafeView Classic (G108) and Safe-Red Gel (G680) can visualize nucleic acids on polyacrylamide gels using a post staining method.
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What device do you use to see the electrophoresis bands? | |
We use both UV, Blue light and LED transilluminators. Our gel QC pictures are taken under UV illumination.
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Will staining appear the same when using UV light and LED (same colors)? | |
UV and LED will give the same colors.
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Which of the SafeView products will work with blue light / LED? | |
All of our SafeView products are compatible with UV and blue light.
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Can SafeView products be stored at -20°C? | |
No, please store SafeView Classic, Safe-Red and Safe-Red Gel at 18-25°C. Safe-Green should be stored at 4°C.
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Can SafeView Classic penetrate cell membranes? | |
Yes, SafeView Classic can penetrate living cell membranes.
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What color is the tracking dye during electrophoresis? | |
Safe-Green and Safe-Red contain a blue tracking dye which will migrate at ~300bp after electrophoresis.
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Can you describe what particular imaging system that one should use to image the gel? | |
Any imaging system with UV, Blue light and/or LED should work.
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