GreenStar™ Nucleic Acid Staining Solution I

Completely dissolve 1 g of agarose in 100 mL 0.5X TBE or 1X TAE Buffer to make a 1% agarose, 1X pre-casting gel. Then, add 500 μl of GreenStar ™ Nucleic Acid Staining Solution I and mix well. Pour into a gel mold and wait until it hardens. After using the gel for electrophoresis, clear bands of nucleic acids can be observed through DUALED Blue/White Transilluminator.

This is because the optimal wavelength of EtBr for excitation is near the UV range while GreenStar™ Nucleic Acid Staining Solution I is near the blue spectrum band. Therefore, stained DNA produces stronger fluorescence under UV transilluminator for the former stain while the latter is more visible under DUALED Blue/White Transilluminator. Nevertheless, if the stained DNA is viewed under the equipments capable of producing light under the solution’s optimal spectrum band, it can be seen that the GreenStar™ Nucleic Acid Staining Solution I can visualize the bands more clearly than EtBr.

• Cause: This may be due to the amphoteric ions included in the GreenStar ™ Nucleic Acid Staining Solution I staining solution. The binding of fluorescence dye with the DNA reduces mobility during the electrophoresis process. This is exceptionally the case when a large amount of nucleic acids are loaded into the gel, as the dye is not completely bound to the samples. With the mixture of bound and unbound samples, the DNA band separation may not be performed well.
• Suggestion 1: As stated in the manual, it is recommended that when being loaded on a pre-cast gel, the DNA samples do not to go over the concentration of 20 ng/band.
  
• Suggestion 2: If DNA samples must go over concentration of 20 ng/band for the electrophoresis, it is highly recommended to use an agarose gel without fluorescent dyes, and then undergo the staining process afterwards. 

The amphoteric ions included in the fluorescent dye of GreenStar™ Nucleic Acid Staining Solution I lower the speed of DNA fragments. The additional time required depends on the size of the agarose gel, but using the solution typically takes about extra 10 minutes than applying EtBr.

Due to the fluorescent dye included in the solution, GreenStar™ Nucleic Acid Staining Solution I may be broken down in proportion to the amount of light irradiation.

If not used, please store at temperature around -20℃ to 4℃ and keep in the dark to protect it against from fluorescent dye degradation.

We recommend to use pre-casting gels made using GreenStar™ Nucleic Acid Staining Solution I within 24 hours.

During the agarose gel preparation process, buffers for its manufacture and the electrophoresis must be the same. If the buffer compositions for the two are different, the concentration of loaded DNA may decrease due to the diffusion process of the ions of the gel and the buffer solution.

The rate of migration of the nucleic acids depends on the ionic strength and the composition of buffer solution. Please choose the buffer according to the characteristics below.

TAE Buffer has lower buffering capacity and ionic strength than TBE buffer, which makes it suitable for undergoing electrophoresis with nucleic acids larger than 12 kb. Also, it can be useful in cloning that uses enzymes for purifying DNA from a gel. However, it must be changed regularly as its long-term usage may induce heat.

On the other hand, TBE buffer has higher buffering capacity and ionic strength than TBE buffer, which makes it suitable for long-term electrophoresis. Also, its high resolution makes it suitable for isolating small nucleic acids smaller than 1 kb. However, TBE buffer contains borate which acts as an enzyme inhibitor, which may interfere DNA purification that requires enzymatic steps.

Completely dissolve 1 g of agarose in 100 mL 0.5X TBE or 1X TAE Buffer to make a 1% agarose, 1X pre-casting gel. Then, add 500 μl of GreenStar ™ Nucleic Acid Staining Solution I and mix well. Pour into a gel mold and wait until it hardens. After using the gel for electrophoresis, clear bands of nucleic acids can be observed through DUALED Blue/White Transilluminator.

This is because the optimal wavelength of EtBr for excitation is near the UV range while GreenStar™ Nucleic Acid Staining Solution I is near the blue spectrum band. Therefore, stained DNA produces stronger fluorescence under UV transilluminator for the former stain while the latter is more visible under DUALED Blue/White Transilluminator. Nevertheless, if the stained DNA is viewed under the equipments capable of producing light under the solution’s optimal spectrum band, it can be seen that the GreenStar™ Nucleic Acid Staining Solution I can visualize the bands more clearly than EtBr.

• Cause: The fluorescence intensity is inversely proportional to the thickness of the manufactured agarose gel: thicker the gel, lower the fluorescence intensity.
• Suggestion 1:: Extend the dyeing time of the gel (10 ~ 40 minutes) or increase the concentration of the staining solution (1X - 10X Recommended, 1/20 - 1/200 Diluted)
  
• Suggestion 2: Use gels with thickness less than 0.5 cm
  
• Suggestion 3: Adjust the exposure time to get the desired image when using Transilluminator for gel photography

Due to the fluorescent dye included in the solution, GreenStar™ Nucleic Acid Staining Solution I may be broken down in proportion to the amount of light irradiation.

When it's not used, please store at temperature around -20℃ to 4℃ and keep in a dark to protect it against from fluorescent dye degradation.

We recommend to use the staining solution diluted with TAE or TBE buffer within 24 hours.

If dyeing performance deteriorates, it is recommended that you use the new staining solution.

During the agarose gel preparation process, buffers for its manufacture and the electrophoresis must be the same. If the buffer compositions for the two are different, the concentration of loaded DNA may decrease due to the diffusion process of the ions of the gel and the buffer solution.

Excessive staining may cause the bands to smear. This is due to the high sensitivity of GreenStar™ Nucleic Acid Staining Solution I. Reduce the dyeing time or the concentration of the staining solution.

Smearing may also occur if the DNA is stained in the degraded state

The rate of migration of the nucleic acids depends on the ionic strength and the composition of buffer solution. Please choose the buffer according to the characteristics below.

TAE Buffer has lower buffering capacity and ionic strength than TBE buffer, which makes it suitable for undergoing electrophoresis with nucleic acids larger than 12 kb. Also, it can be useful in cloning that uses enzymes for purifying DNA from a gel. However, it must be changed regularly as its long-term usage may induce heat.

On the other hand, TBE buffer has higher buffering capacity and ionic strength than TBE buffer, which makes it suitable for long-term electrophoresis. Also, its high resolution makes it suitable for isolating small nucleic acids smaller than 1 kb. However, TBE buffer contains borate which acts as an enzyme inhibitor, which may interfere DNA purification that requires enzymatic steps.