ViSafe Green Gel Stain is a stable, sensitive and environmentally safe fluorescent nucleic acid dye for staining double-stranded DNA (dsDNA), single-s tranded DNA (ssDNA) or RNA in agarose gels or polyacrylamide gels.

ViSafe Green Gel Stain has UV absorption between 250nm and 300nm. It is compatible with either 254nm UV transilluminator or a gel reader equipped with visible excitation. Both green gel stain and EtBr have the same spectra, so the green gel stain replaces the highly toxic Ethidium Bromide (EtBr) without changing existing imaging system.

The dye is confirmed by Ames test results that it is impenetrable to latex gloves and cell membranes. By using the suggested working concentrations in gel staining, the dye is proven unable to cross cell membranes; and it is noncytotoxic and nonmutagenic at working concentrations.

ViSafe Green Gel Stain, 10000X in H2O, can be diluted 10000X for precast gel protocol or 3000-3300X for post gel staining. One vial (0.5ml) of 10000X solution can be used for at least 100 minigels either using precast method or post-staining method.

Stable to be microwaved or being heated. The working solution is stable at room temperature when kept in dark. Wide application Suitable to stain dsDNA, ssDNA and RNA.

It is suitable to use in agarose gel or polyacrylamide gel and compatible with downstream applications, such as gel recovery & cloning.

Easy staining protocols and easy precast gel staining & post-staining procedures.

Compatible with most imaging system Gel can be viewed with standard UV transilluminator, visible light gel reader, or other gel imaging system.

Shipment: @ ambient temperature

Storage: 4-8°C or -20°C

Precast Protocol for 1% Agarose Gel

1. Pour 1g of agarose powder and 100ml 1X TAE or 1X TBE into glass
    flask.

2. Melt the agarose in microwave for 1-3 mins until the agarose is
    completely dissolved.

3. Add 3-5µl GreenGel stain per 100ml gel and stir the gel solution to mix
    thoroughly.

Make sure the red gel stain is swirled and stirred well to mix with gel solution.

4. Pour the agarose solution onto gel plate and insert a comb.

5. Place newly poured gel at 4°C for 10-15 mins or stay at room
    temperature for 20-30 mins, until it has completely solidified.

*Extra agarose containing the red gel stain can be kept in solid form at 4°C and can be remelted to cast more gels.

6. After the gel is ready, perform gel electrophoresis.

7. Visualize or image the gel directly under UV light or blue light after gel electrophoresis is done. Standard transilluminator or ethidium bromide filter can be used for gel imaging purpose.

Recomendations:

*Dilute the stock solution into agarose gel solution at 1:10000.

*Since the red gel stain is thermally stable, the stock solution can be added while the gel solution is still hot.

*The greengel stain can be pre-combined with agarose powder and gel working solution followed by microwaving or other heating procedures.

Remark:

- However, more dye might be necessary to be added for
  optimal signal.

- ViSafe Green Gel Stain precast gels can be prepared in
  larger quantities and stored for later use since the stain is
  hydrolytically stable. Any leftover gel solution can be stored
  and re-heated for additional gel casting. Avoid heating the
  green gel stain gel solution more than 3 times.  

Remark:

Due to ViSafe Green Gel Stain’s slow diffusion rate in the relatively tight polyacrylamide gel matrix, the green gel stain is not suggested for staining DNA or RNA in precast polyacrylamide gels. Post-staining method can be used for polyacrylamide gels.

Using precast gel with green gel stain is more convenient. However, some DNA samples may experience migration retardation or compromised resolution in the presence of green gel stain. The green gel stain may affect DNA migration during electrophoresis. Hence, post-staining with green gel stain is highly recommended. Post-staining method may lead to gel results with higher sensitivity and without dye interference with DNA migration. Besides, post-staining method is a simple protocol with no destaining and no special buffer needed.

Post-staining protocol for Agarose Gel and Polyacrylamide Gel

1. Run the gel electrophoresis accordingly.

2. Dilute 10000X ViSafe Red Gel Stain stock solutions to 3X staining solutions with 0.1M NaCl water solution. (This solution can be used at least 2-3 times, protected from light. Suggested to use container covered with alumin ium foil or use dark colour container.)

Example: 30µl of red gel stain added into 100ml of 0.1 M NaCl solution

*NaCl solution in the staining solution is optional. Adding NaCl in the staining solution enhances the staining, but may promote dye precipitation if the staining solution is to be used repeatedly. Any staining solution to be reuse d is preferably stored at room temperature in a dark pla ce to reduce possible dye precipitation problem.

3. Remove the gel from the gel tank and transfer into staining container.

4. Allow the gel to stain for at least 25-30 mins with gentle shaking.

5. Destaining time may vary depending on the thickness of the gel and percentage of agarose.

6. Visualize or image the stained gel under UV light or blue light using standard transilluminator or ethidium bromide filter.

Recomendations:

1. To increase resolution of the DNA bands:

a. Running the gel at a lower voltage for longer period of time

b. Using a wider gel comb

c. Loading less DNA into well

2. To get better separation of bands, adjust the agarose percentage of the gel if the similarly sized bands that that are running too close together are loaded. A higher percentage agarose gel will help in resolving smaller bands from each other, and a lower percentage gel will help in separating larger bands.

3. ViSafe Green Gel Stain is efficiently removed from DNA by phenol/chloroform extraction and ethanol precipitation.

1. To increase resolution of the DNA bands:

a. Running the gel at a lower voltage for longer period of time

b. Using a wider gel comb

c. Loading less DNA into well

2. To get better separation of bands, adjust the agarose percentage of the gel if the similarly sized bands that bands that are running too close together are loaded. A higher percentage agarose gel will help resolving smaller bands from each other, and a lower percentage gel will help separating larger bands.

3. ViSafe Red Gel Stain is efficiently removed from DNA by phenol/chloroform extraction and ethanol precipitation.