Size: 500 µl in water.
GRGreen is a nucleic acid stain for detecting nucleic acids, e.g., double-stranded DNA, in agarose gel. It can be used for replacing mutagenic ethidium bromide (EB).
Compared to EB which is a very strong mutagen, GRGreen caused fewer mutations than EB in the Ames test.
- Available at 10,000X in H2O for better safety.
- Compatible with UV or blue light transilluminator and common gel documentation systems.
- Will not affect downstream experiments: compatible with all gel purification kits tested, will not inhibit ligation reaction etc.
- Compatible with Sodium Borate Electrophoresis Buffer: Run gel 2-3 times faster at higher voltage, resolve shaper bands in minutes, and less heat generation.
- Cut out DNA bands for subclonning under safer blue light: No mutations caused by EB and UV light.
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Fig. 1, Sensitivity of GRGreen stain.
1% agarose gel prepared with SeaKem LE Agarose (Lonza) and 1X GRGreen. 75V, 0.5X TAE.
Lefe: 100 bp DNA ladder, 250ng, 125 ng, 62.5 ng and 31.3 ng, total ng DNA per lane.
Right: 1kb DNA ladder, 250ng, 125 ng, 62.5 ng and 31.3 ng, total ng DNA per lane.
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| Stain A |
GRGreen |
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| DarkReader Blue Light Transilluminator |
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| Stain A |
GRGreen |
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| UV Transilluminator |
Fig. 2, GRGreen in water is more sensitive than Stain A in water.
1% agarose gel prepared with SeaKem LE Agarose (Lonza) and 1X GRGreen or 1X Stain A (from competitor A), run with 1X Lithium Borate Fast Electrophoresis Buffer at 100 V.
Left: 100 bp DNA ladder, 250ng, 125 ng, 62.5 ng and 31.3 ng, total ng DNA per lane.
Right: 1kb DNA ladder, 250ng, 125 ng, 62.5 ng and 31.3 ng, total ng DNA per lane.
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| TAE |
TBE
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Sodium Borate |
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Fig. 3, GRGreen is compatible with three common running buffers.
1% agarose gel prepared with SeaKem LE Agarose (Lonza) and 1X GRGreen, run with 1X three Electrophoresis Buffers at 125 V.
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| 100 bp |
1kb Plus |
1kb |
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Fig. 4, Lithium Borate Fast Electrophoresis Buffer gives the most satisfactory resolution.
1% agarose gel prepared with SeaKem LE Agarose (Lonza) and 1X GRGreen, run with 1X Lithium Borate Fast Electrophoresis Buffer at 100 V.
Storage: Store at 4 oC.
Protocol:
1. Prepare 20 to 40 ml of agarose gel solution (concentration from 0.7~2.0%) with TAE, TBE or Borate Buffer in a 250 ml flask and mix it thoroughly. Place the flask in the microware, heat on high until the solution is completely clear and no small floating particles are visible (about 2~3 minutes).
2. After the gel solution cool to about 55 oC, add GRGreen to the solution to 1X final concentration. Swirl the flask gently to mix the solution and avoid forming bubbles.
3. Pour the gel solution into a gel tray until the comb teeth are immersed about 1/4~1/2 into the gel solution.
4. After the agarose gel has solidified you can perform electrophoresis using either 0.5 to 1X TAE, TBE or Borate Buffer.
5. Detect the bands using UV or blue light transilluminator.
FAQ
1, Should I wear gloves when using this dye?
You should exercise common safe laboratory practices when using this reagent.
2, What blue light transilluminator should I used with GRGreen dye?
DarkReader.
3, What filter should I use for blue light transilluminator?
Filter from Clare Chemical.
MSDS: GRGreen MSDS, Disposal Guide
Protocol
