GRGreen Nucleic Acid Stain, 10,000X in DMSO
Size: 500 µl
Description GRGreen is a nucleic acid stain for detecting double-stranded DNA, single-stranded DNA, and RNA in agarose gel. It can be used for replacing mutagenic ethidium bromide (EB). This nucleic acid stain has two fluorescence excitation maxima when bound to nucleic acids, one centered at approximately 300 nm and one at approximately 500nm. Thus, GRGreen is compatible with either a UV transilluminator or a blue light transilluminator.
Features
- Stains RNA, DNA and oligonucleotides. Can stain DNA ranging from 10 bp to 100kb.
- Ultrasensitive: detect DNA at picogram level.
- DNA bands can be viewed using either UV or safer blue light transilluminator. If you use a blue light transilluminator, you will not expose to dangerous UV light, so you will not get sun burn or skin cancer or damaged eyes.
- You can use digital camera to take gel photos: Our gel documentation system is recommeded.
- Will not affect downstream experiments: Compatible with all gel purification kits tested, will not inhibit ligation reaction etc.
- Cut out DNA bands for subclonning under safer blue light: No mutations caused by EtBr and UV light.
- Watch DNA migrate on your bench with a blue light transilluminator, in real-time without UV light.
Sensitivity
Detection limits of common nucleic acid stains using Clare Chemical's DarkReader
| Stain |
Visual Detection Limit
(pg dsDNA)
|
Detection Limit using Camera
(pg dsDNA) |
| GRGreen |
100 |
10 |
| GelStar |
100 |
20 |
| GelGreen |
250 |
40 |
| SYBR Safe |
1500 |
100 |
The following pictures show that GRGreen is more sensitive than SYBR Safe and GelGreen (this gel photo was selected from a few similar comparison experiments provided by independent laboratories).
Safety
Cytotoxicity Test
CEM cells were cultured in RPMI 1640 with 10% FBS. Blindly coded SYBR Safe and GRGreen were added to the cells at 1 X final concentration and the cells were cultured at 37oC in a CO2 incubator. Cells were collected at different time points, and the cytotoxicity was measured using a CCK-8 kit purchased from Dojindo Molecular Technologies (Gaithersburg, Maryland, USA). This test was done by an independent laboratory in Maryland, USA.
Result: GRGreen is less cytotoxic than SYBR Safe.
Storage: 4 oC or -20 oC for long term storage.
Disposal: as regular trash.
Staining Protocols
Post-Staining Protocol
Prepare enough 1X staining solution, e.g., add 2~5 µl of 10000X GRGreen Stain to 20~50 ml TE, TAE, TBE 0.1M NaCl or water. After electrophoresis, place the gel in a polypropylene tray and add staining solution to cover the top of the gel. Agitate the gel gently at room temperature using a rocking shaker while keeping the solution away from bright light. The staining process (at room temperature) is about 15-30 minutes depending on the thickness of the gel. View the gel using a UV or blue light transilluminator.
In-Gel Staining Protocol
Prepare the gel solution and cool the agarose solution to 55~65 °C. Add 10000X GRGreen stain to the gel to final concentration of 1X, for example, add 5 µl 10000X GRGreen stain to 50 ml gel solution. Mix the gel solution thoroughly then immediately cast the gel and allow the gel to solidify. Load samples and run the gels using your standard protocol. View the gel using a UV or blue light transilluminator. Do not overload DNA (optimal amount of DNA should be at least 10 times less than what you use with EtBr).
Double-Staining Protocol
1, Preparation of 6X GRGreen Loading Buffer
Add 1 µl of 10000X GRGreen to 100 µl of 6X Loading Dye and mix. The resulting buffer is called 6X GRGreen Loading Buffer. This buffer is stable at 4 °C for a few days.
2, Pre-Staining: Mix your DNA sample with 6X GRGreen Loading Buffer. For example, add 2 µl of GRGreen Loading Buffer to 10 µl of your DNA sample, mix. optimal amount of DNA should be at least 10 times less than what you use with EtBr.
3, Prepare the gel solution and cool the agarose solution to 55~65 °C. Add 10000X GRGreen stain to the gel to final concentration of 1X, for example, add 5 µl 10000X GRGreen stain to 50 ml gel solution. Mix the gel solution thoroughly then immediately cast the gel and allow the gel to solidify.
4, Load 2 to 12 µl of your “pre-stained” samples and run the gels using your standard protocol. View the gel using a UV or blue light transilluminator.
In-Gel Staining Double-Staining
Other Applications
GRGreen may be used for:
- Quantitation of DNA
- Nucleic acid detection, e.g., isothermal nucleic acid amplification such as RCA and loop-mediated isothermal amplification.
Protocol: GRGreen DNA Stain Protocol
MSDS: GRGreen MSDS, Disposal Guide
