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RedSafe™ Nucleic Acid Staining Solution (20,000x) is a new and safe nucleic acid staining solution.
It is an alternative to the traditional ethidium bromide(EtBr) stain for detecting nucleic acid in agarose gels. It emits green fluorescence when bound to DNA or RNA. This new stain has two fluorescence excitation maxima when bound to nucleic acid, 309 nm and 419 nm. It has a visible excitation at 514 nm. The fluorescence emission of RedSafe™ bound to DNA is at 537 nm. RedSafeTM Nucleic Acid Staining Solution (20,000x) is as sensitive as EtBr. The protocol is also similar with a protocol of EtBr. The Ames test proves lower mutation than RedSafe™ Nucleic Acid Staining solution; Comparable to EtBr. RedSafe™ Nucleic Acid Staining Solution (20,000x) has a negative result in mouse marrow chromophilous erythrocyte micronucleus tests and mouse spermary spermatocyte chromosomal aberration tests.
Therefore, RedSafe™ Nucleic acid Stainig Solution (20,000x) is superior to EtBr for detecting nucleic acid in agarose gels.
APPLICATIONS
♦ Visualization of DNA and RNA bands as they separate during agarose gel electrophoresis
♦ Isolation of DNA fragments for subcloning without mutations normally caused by EtBr
KIT CONTENTS
♦ RedSafe™ Nucleic Acid Staining Solution 1 ml
STORAGE
♦ Room Temperature
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 RedSafeTM 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 RedSafeTM 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.
5. Detect the bands under UV illumination.
Note : RedSafeTM 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.
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