RedSafe Nucleic Acid Staining Solution (20,000x) is a new and safe nucleic acid staining reagent, a substitute for EtBr (ethidium bromide)
The RedSafe Nucleic Acid Staining Solution exhibits two fluorescence excitation maxima at 309 nm and 419 nm when bound to nucleic acids and is visible at a wavelength of 514 nm. Red Safe Nucleic Acid Staining Solution is as sensitive as EtBr and its usage is similar to EtBr. Compared with EtBr, a potent carcinogen, Red Safe Nucleic Acid Staining Solution showed almost no mutation in the Ames test and negative results in mouse marrow chromophilous erythrocyte micronucleus test and mouse spermary spermatocyte chromosomal aberration test. Therefore, we strongly recommend that you select RedSafe™ Nucleic Acid Stainig Solution (20,000x) instead of EtBr to identify nucleic acids on agarose gel.
PROTOCOL
1. Prepare a 100 ml of
agarose gel solution (concentration from 0.8~3 %) in a 250 ml flask and mix it thoroughly. Place the flask in the microwave, heat in until the solution is completely clear (about 2~3 minutes). Note : The thickness of gel should be less than 0.5 cm since thick gels may decrease sensitivity.
2. Add 5 ㎕ of RedSafe Nucleic Acid Staining Solution (20,000x) to the agarose solution. Swirl the flask gently to mix the solution and avoid forming bubbles.
3. While the agarose solution cools, pour it into the gel tray until the comb teeth are immersed about 1/4~1/2 into the agarose. Note : Repeated melting of gels containing RedSafe Nucleic Acid Staining Solution (20,000x) may result in low sensitivity.
4. Allow the agarose gel to cool until solidified. Load samples on the gel and perform electrphoresis.
Note : RedSafe Nucleic Acid Staining Solution (20,000x) allows visualization of DNA (>50 ng) in the agarose gel under visible light. This eliminates the need for exposure to UV light, which may nick and damage DNA. The intact DNA fragments purified from agarose gel can increase the efficiency of subsequent molecular biology manipulations such as cloning, transformation and transcription.