SYBR Safe DNA Gel Stain

Staining nucleic acids after electrophoresis

Soak the gel in SYBR Safe stain. If using SYBR Safe gel stain concentrate, dilute 10,000X in TAE or TBE buffer (as appropriate) prior to use. Place the gel in a plastic container, such as a pipet-tip box lid or a household food-storage container. Do not use a glass container, because the dye in the staining solution may adsorb to the walls of the container, resulting in poor gel staining. Add sufficient SYBR™ Safe DNA gel stain to cover the gel. A 50 mL volume is sufficient for staining most standard minigels. To stain larger gels, increase the volume of staining solution in proportion to the increased gel volume, and ensure that the gel is fully immersed during staining.
Incubate for 30 minutes. Cover the gel and the staining solution with aluminum foil or place them in the dark to protect from light. Continuously agitate the gel on an orbital shaker at 50 rpm. No destaining is required.

Precasting SYBR Safe Stain in agarose gels

Prepare the agarose gel directly in SYBR Safe DNA gel stain. SYBR Safe stain is provided in buffer; simply substitute SYBR Safe stain for the buffer when preparing the molten agarose. If using the 10,000X SYBR Safe stain concentrate, dilute the concentrated stain 1:10,000 in agarose gel buffer (e.g., 1X TBE or 1X TAE) and add the buffer plus stain mixture to the powdered agarose. For example if you run TBE gels and require 30 mL of molten agarose for your tray, add 3 μL of 10,000X SYBR Safe stain concentrate to 30 mL of 1X TBE, mix well, and add to the powdered agarose.
Note: You can heat the agarose/SYBR Safe stain mixture in the microwave. As with precasting gels with ethidium bromide, the mobility of nucleic acid fragments in the gel may be somewhat slower when run in these gels compared to their mobility in the gel without stain.
Run the gel. Use a running buffer appropriate to the SYBR Safe gel stain formulation. No post-staining or destaining is needed.

Viewing and photographing the gel

You can view stained gels using a standard 300 nm transilluminator, a 254 nm epi- or transilluminator, or a blue-light transilluminator such as the Safe Imager 2.0 Blue-Light transilluminator. DNA stained with SYBR Safe stain can also be visualized and analyzed using imaging systems equipped with an excitation source in the UV range or between 470–530 nm. 
Note: If bands from the SYBR Safe stained gel are to be excised and used in a ligation reaction, we recommend that the gel is illuminated using blue-light source (i.e., Safe Imager 2.0 Blue-Light transilluminator) and not a UV light source. In some instances, UV light sources in combination with SYBR Safe stain can lead to reduced cloning efficiencies.
You can photograph stained gels using Polaroid 667 black-and-white print film and SYBR Safe photographic filter. SYPRO photographic filter or a Kodak Wratten #9 filter also work well. Using this film and one of these filters, SYBR Safe DNA gel stain provides the same detection sensitivity as ethidium bromide and an appropriate filter. Do not use a standard ethidium bromide photographic filter with SYBR Safe DNA gel stain. Gels stained with SYBR Safe stain can also be imaged using a CCD camera or a laser-based scanner.