Helixyte™ Green Nucleic Acid Gel Stain *10,000X DMSO Solution*

Spectral Properties of Helixyte™ Green Nucleic Acid Gel Stain

Excitation/Emission: 497/521 nm when bound to DNA

Helixyte™ Green nucleic acid gel stain is significantly less mutagenic than ethidium bromide. Nevertheless, we must caution that there is currently no available data on the mutagenicity or toxicity of Helixyte™ Green stain in humans. It is prudent to treat this reagent as a potential mutagen and handle it with appropriate care, especially considering its binding affinity to nucleic acids. Additionally, ensure proper disposal of the stain in compliance with local regulations.

We have observed that the greatest sensitivity is attained through post-staining, which eliminates the risk of dye interference with DNA migration. While the precast protocol is more convenient, some DNA samples may experience migration issues. Hence, it is strongly advised to limit the gel running time to no more than 2 hours. The provided protocols are recommendations, and conducting comparisons may be necessary to determine which one better meets your specific needs.

1. Prepare a 1X Helixyte™ Green working solution by diluting the 10,000X stock reagent with a pH 7.5 - 8 buffer, such as TAE, TBE, or TE. Buffers with a pH of 8.0 are preferred.

   Note: Staining solutions prepared in water are less stable than those prepared in buffer and must be used within 24 hours to ensure maximal staining sensitivity.

   Note: In addition, staining solutions prepared in buffers with pH below 7.5 or above 8.0 are less stable and show reduced staining efficacy.

1. Run gels based on your standard protocol.

2. Place the gel in a suitable polypropylene container. Gently add a sufficient amount of the 1X Helixyte™ Green working solution to submerge the gel.

   Note: Do not use a glass container, as it will adsorb much of the dye in the staining solution.

3. Agitate the gel gently at room temperature for ~30 minutes, protected from the light.

   Note: The staining solution can be stored in the dark (preferably refrigerated) for a week and reused up to 2 - 3 times.

4. Image the stained gel with a 254 nm transilluminator or a laser-based gel scanner using a long path green filter, such as a SYBR® filter or GelStar® filter.

1. Prepare agarose gel solution using your standard protocol.

2. Add 1X Helixyte™ Green working solution to the gel and mix thoroughly.

3. Run gels based on your standard protocol.

4. Image the stained gel with a 254 nm transilluminator or a laser-based gel scanner using a long path green filter, such as a SYBR® filter or GelStar® filter.

1. Incubate DNA with a 1:3000 to 1:10,000 dilution of the dye (in TE, TBE, or TAE) for at least 15 minutes prior to electrophoresis.

2. Run gels based on your standard protocol.

3. Image the stained gel with a 254 nm transilluminator, or a laser-based gel scanner using a long path green filter such as a SYBR® filter or GelStar® filter.