|Description||Safe-Green™ is a new and safe nucleic acid stain for the visualization of nucleic acids in agarose and polyacrylamide gels. This dye eliminates the need for toxic Ethidium Bromide (EtBr, a potent mutagen), commonly used in gel electrophoresis.
|SafeView Series||Loading Dye|
|LED Viewer Compatibility||Yes|
|Applications||Safe Detection of dsDNA, ssDNA and RNA in agarose and polyacrylamide gels.|
|Note||All SafeView DNA Stains are ISO-13485 certified.
Dispose of SafeView DNA Stains as you would any other non-carcinogenic fluorescent dye (eg. Acridine orange; Propidium iodide).
|Shipping Conditions||Shipped on blue ice packs.|
|Storage Condition||Store at 4°C for up to 2 years.|
|Unit quantity||1.0 ml|
Can SafeView be a replacement for ethidium bromide? Can I do gel extraction with it?
SafeView can be used as a replacement for ethidium bromide as they both work on general agarose. We recommend using SafeGreen for downstream cloning applications as SafeView can interfere with the ligation reaction, yielding fewer colonies.
How does SafeView work and why is it not carcinogenic?
There are fluorescent compounds in SafeView and these fluorescent compounds have the capability to bind to DNA. There may be some unknown effects of SafeView that have not been documented but that applies to the SYBR set as well; however, SafeView products are definitely not as carcinogenic as ethidium bromide.
How do I use SafeView products?
The Safe-(Red, Green and White) loading dyes work the same as 6x loading dye, loaded with the sample. SafeView Classic is used directly in the gel and the running buffer.
Does the SafeView differentiate double stranded nucleic acid and single stranded nucleic acid? Does the Safe-(Red, Green and White) work the same way?
Under UV, SafeView Classic emits a green fluorescence when bound to both single and double stranded DNA templates, therefore they cannot be differentiated by this method. It will emit a red fluorescence when bound to RNA templates. The SafeView Stains (Red, Green and White) do not perform in this way and will stain all nucleic acid templates one color.
At what temperature do I store the SafeView products?
All the SafeView products should be stored at four degrees Celsius.
Do I need a special filter for photography of DNA gels stained with SafeView?
Under UV light, SafeView Classic emits a green fluroescence when bound to both single and double stranded DNA templates. It will emit a red fluorescence when bound to RNA templates. No filter is necessary for viewing these colours, however a filter may be needed for photographing the gel.
How long does the SafeView Classic stain last in a gel?
Our in-house testing has shown that SafeView 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, at 4C in the dark, sealed in a plastic bag or pouch with wet paper towel loosely wrapped around the gel.
Why is SafeView (G-108) stain not working on my samples?
Make sure you are following the protocol carefully. It is critical that both buffer and gel have SafeView dye in them otherwise it will not work. This is different from ethidium bromide. You can consider to add 2.5ul of SafeView (instead of 5ul) for every 100ml of running buffer, which will reduce background fluorescence and allow the bands to show with more contrast.
Is it degradable, if so how fast and under what circumstances?
2 hours over 100C
What is the sensitivity of the dyes?
Safe-Green has Excitation Wavelength of 490nm and Emission Wavelength of 525nm, and its sensitivity range is between 0.2-0.6ng. Safe-Red has Excitation Wavelength of 540nm and Emission Wavelength of 630nm, and its sensitivity range is between 0.3-0.8ng. Safe-White has Excitation Wavelength of 370nm and Emission Wavelength of 470nm, and its sensitivity range is between 0.2-0.5ng. SafeView Classic emits green fluorescence when bound to dsDNA and ssDNA and red fluorescence when bound to RNA. This stain has one excitation (490 nm) and two emission spectra (520 nm and 635 nm) and the sensitivity range is between 0.1-0.3ng. SafeView Plus has Excitation Wavelength of 490nm and Emission Wavelength of 525nm and its sensitivity range is between 0.05-0.1ng.
Can SafeView products be used post-stain?
Only SafeView Plus (G468) should be used in a post stain. SafeView classic and Safe Stains are not designed for post-staining. SafeView Classic must be added to the gel and the running buffer prior to the loading of the samples. Safe-(Red, Green and White) stains must be added to the sample before loading it to the gel.
Why is the EtBr signal stronger in the pictures when I compare SafeView with EtBr?
A reason for this is that most gel doc systems have been optimized for EtBr so that is why the EtBr signal may be stronger in the pictures.
What is the functional pH range for SafeGreen?
SafeGreen works best between pH 7 to 9.
will I need an additional loading buffer for my samples?
The loading dye is included in the products. No additional loading buffer needed.
What is the difference between Safe Green, Safe Red and Safe White? Does the loading buffer have different colors? Which filters do I have to use to detect light emission?
The difference is simply in colour of the dye, when viewed under UV the bands in the gel will show up in green, red or white.
What color is the dye during electrophoresis?
The color of the dyes seen during migration would be blue, the same for all 3 dyes (Safe-Green/ Safe-Red/Safe-White). The size of the dye should be at approximately 300bp after electrophoresis. All three loading dyes have different excitation and emission wavelengths, so they will emit different colors. When bound to DNA, Safe-Green emits green fluorescence, Safe-Red emits red fluorescence, and Safe-White emits whitish pale blue fluorescence.
We see migrations and band shifting of our fragments. Are there any recommendations that you can give us to minimize this band shifting?
Shifting is unavoidable and quite natural for any fragments, regardless of the staining agent. We suggest to use SafeGreen ladders, which will give accurate molecular weight with no additional staining agents needed. http://www.abmgood.com/DNA-Ladder-Safe-Green™-100bp-Opti-DNA-Marker-Invitroge-G473.html http://www.abmgood.com/DNA-Ladder-Safe-Green™-1kb-Opti-DNA-Marker-Invitroge-G474.html
We see shifting and migration of the DNA fragments. What are the recommendations to minimize this?
Shifting is unavoidable and quite natural for such fragments, regardless of the staining. We suggest using SafeGreen ladders, which will give accurate molecular weight with no additional staining agents needed. http://www.abmgood.com/DNA-Ladder-Safe-Green™-100bp-Opti-DNA-Marker-Invitroge-G473.html http://www.abmgood.com/DNA-Ladder-Safe-Green™-1kb-Opti-DNA-Marker-Invitroge-G474.html
I cannot see 100bp and 200bp bands on a 1% gel. What should I do?
It is very difficult to detect 100bp and 200bp bands in 1% gel with any stains. Higher gel concentrations should be used, such as 2% agarose.
Can I use SafeView products in Polyacrylamide gels?
Yes, we have tested our SafeView products for this application.
Which of the Safe stains will work with blue light / LED?
All of our SafeView stains have been tested in-house to be compatible with UV light. SafeView Classic, SafeView Plus, and SafeGreen will also work under blue light/LED. SafeRed and SafeWhite will only work under UV light. Safe View Plus should only appear green. SafeView Classic will be red for RNA and green for DNA.
Are SafeGreen stained DNA bands suitable for ligation purposes?
Does the dye migrate in a 1% agarose gel?
Yes, the dye should migrate on a 1% gel.
Does G108-G contain SDS?
What is the concentration of G108-G?
6X loading dye
Would adding Safe-Green to the DNA ladder interfere with the migration rate of the loading dye of the ladder?
No, adding SafeGreen to the DNA ladder will not interfere with the DNA ladder’s loading dye. You still need to add SafeGreen to the DNA ladder at the same ratio. To be more technically correct, SafeGreen, and all other EB alternative DNA binding dyes, would alter the migration pattern of DNA on agarose gel by shifting DNA to a larger molecular size; it is just simply because the overall molecular size of a particular DNA band increases as more dye is binding onto it. This is why we recommend mixing the DNA ladder and SafeGreen at the exact same 1:6 ratio, so everything on the gel will be subjected to the same concentration of SafeGreen and thus having the same shift.
What is the size of the dye in SafeGreen after electrophoresis?
The dye should be at ~300bp after electrophoresis.
Can Safe-Green penetrate the nucleus?
Yes, our Safe-Green™ stain indeed has the ability to penetrate the nucleus.