DyeNA-View™ is a new nucleic acid stain, a safe alternative to the traditional ethidium bromide (EtBr) stain for detecting nucleic acids in agarose gels. It emits green fluorescence when bound to DNA or RNA. This new stain has two fluorescence excitation peaks when bound to nucleic acid: one at 267nm and another at 294nm. In addition, it has one visible excitation peak at 491nm. The fluorescence emission of DyeNA-View™ bound to DNA is centered at 530nm.

Storage

Store at 4 °C for 2 years.

Protocol

1. Prepare 100mL of agarose gel solution (concentration from 0.8-2%) in a 250mL flask and mix thoroughly.

 Place the flask in a microwave oven, heat until the solution is completely clear and no small floating particles are visible (about 2-3 minutes).

2. Let agarose solution cool down to about 50 °C (about when you can comfortably keep your hand on the flask), about 5 mins.

3. Vortex the DyeNA-View™ tube for 2-3 seconds. Add 2-5µL of DyeNA-View™ to the gel solution. Swirl the flask gently to mix the stain solution into the molten agarose while avoiding bubbles.

4. Pour enough gel solution into the gel tray until the comb teeth are immersed at least about ¼-½ into the gel solution.

5. Allow the agarose gel to cool until solidified. Load samples on the gel and perform electrophoresis.

[Optional: For increased sensitivity, after gel is electrophoresed, perform a post stain rinse by incubating the gel in distilled water for 5 min.]

6. View the DNA bands under UV/Blue LED illumination.

Notes

1. The thickness of the gel should be less than 0.5cm since thick gels may decrease the sensitivity of DNA detection.

2. Repeated melting of the gels containing DyeNA-View™ may result in low sensitivity of detection.

3. The intact DNA fragments purified from agarose gel stained with DyeNA-View™ can be used in subsequent molecular biology applications such as cloning, transformation and in vitro transcription.

4. DyeNA-View™ may irritate skin and eyes. Please wear gloves while handling it.

Typical Results

See picture-A commercial 100bp ladder was loaded in 10µL and 5µL volume into each of two wells. RedSafe™ (Froggabio) and DyeNA-View™ (GeneBio Systems, Inc.) were used according to instructions provided by the manufacturers.