GREEN STAIN

GREEN STAIN: FOR DNA DETECTION ON AGAROSE GELS
Nucleic acid gel staining has been performed with ethidium bromide for decades. Even though ethidium bromide is very economic and easy to use, its mutagenicity and hazardous waste disposal fueled the development of DNA gel staining alternatives. Cyanagen has developed Green Stain to solve these issues. Green Stain has a high binding affinity for nucleic acids providing bright green light-emitting DNA-dye complexes. Bright green fluorescent bands and very low background fluorescence are its major features. In addition, Green Stain has a very low toxicity combined with an extraordinary sensitivity. It can be used for any gel staining application requiring the detection of up to 100 pg of genomic dsDNA with sufficient sensitivity. Green Stain preferentially binds double-strand DNA, it also allows also single-strand DNA and RNA detection, although with lower sensitivity.
Features:

Very low toxicity;

• Ready to use kit;

• High sensitivity: detection up to 100 pg per band;

• Optimal signal to background ratio;

• Simple substitution of your stain reagent with Green Stain

Staining with GREEN STAIN Kit components

• One package includes 1 x 500µl Green Stain concentrate in DMSO.

Other materials required

• TE (10 mM Tris-HCl, 1 mM EDTA, pH 7.5), TBE (89 mM Tris base, 89 mM boric acid, 1 mM EDTA, pH 8) or TAE (40 mM Trisacetate, 1 mM EDTA, pH 8) buffer

• Agarose

Procedures

Green Stain can be used with pre-, post- electrophoresis gel staining and DNA sample pre-staining.

Pre-electrophoresis gel staining:

  • Dissolve the agarose in the buffer using microwave or heating appliance.
  • Thaw Green Stain at room temperature
  • Add Green Stain to the gel solution (1:10000 dilution). Mix thoroughly.
  • Pour the gel and let it cool down.
  • Perform electrophoresis in an agarose gel in TBE or TAE buffers according to standard procedures.
  •  Visualize or document the gel
Sample pre-staining:

  • Dilute Green Stain 1:100 in TE or TBE buffer.
  • Use the Green Stain working solution as 2X and add it to the DNA sample plus loading dye and incubate at least 5 min.
  • Perform electrophoresis in TBE or TAE buffers according to standard procedures.
Post-electrophoresis gel staining:

  • Perform electrophoresis in TBE or TAE buffers according to standard procedures.
  • Thaw Green Stain at room temperature and dilute 1:2000 in TE, TBE, or TAE buffers.
  • Place the gel in the staining solution and incubate at room temperature for 15–40 minutes with gentle agitation. Always wrap the staining container in aluminum foil to protect from direct light.
  • Visualize or document the gel
Imaging: The nucleic acid-bound Green Stain is efficiently excited at ~254 nm and ~488 nm. Detection can be performed with the same instruments used for ethidium bromide and SYBR® Green gel staining such as standard UV transilluminator (254 nm) as well as with CCD-camera imaging system or laser-based scanner selecting the SYBR® Green filter.
TWO VARIANTS GREEN STAIN GREEN STAIN WITH LOADING DYE