Description:
Gel RedSave fluorescent Stain (10000X) is a highly sensitive and environmentally safe fluorescent nucleic acid dye for detection of DNA and RNA in agarose and polyacrylamide gels.
Gel RedSave fluorescent Stain emits red fluorescence when bound to dsDNA, ssDNA, and RNA and replaces the need for the mutagenic ethidium bromide, commonly used in gels staining procedures for visualization of nucleic acids.
Gel RedSave fluorescent Stain is not cell-permeable and non-mutagenic in the Ames test.
DNA visualization after staining with Gel RedSave fluorescent Stain can be done using conventional gel imaging instruments (UV transilluminators).
The blue LED illumination can be used too, however, Gel RedSave fluorescent Stain stained nucleic acid bands will be less intensive.
For blue LED transilluminators, we recommend our Gel GreenSafer fluorescent Stain.
The nucleic acid dye is efficiently removed from DNA by gel extraction or ethanol precipitation, and therefore does not interfere with downstream DNA manipulations such as restriction digestion, ligation, PCR, and sequencing.
Gel RedSave fluorescent Nucleic Acid Safe Gel Stain, 10,000X stock is a concentrated solution that can be diluted 10,000 times for use in precast gel staining or 5,000 times for use in post gel staining according to the procedures described in the product manual.
Features:
· Detection of dsDNA, ssDNA and RNA in gel by pre-staining or post-staining
· Safer alternative to the ethidium bromide staining
· Highly sensitive (0.6 ng for 500 bp DNA fragment)
· Compatible with various gel imaging instruments
Shipment: @ ambient temperature
Storage: 4-8°C or room temperature
Final price excl. shipping costs3
Figure 1. Comparison of nucleic acid gel stains. Serial dilutions of 1kb DNA Ladder ranging from 10 to 30 ng total DNA per lane were run on 1 % agarose gel precast with SYBR® Safe DNA Gel Stain (left panel) and Gel RedSave fluorescent Stain Nucleic Acid Safe Gel Stain (right panel). Bands were visualized by exposure to UV light (312 nm).
1. To increase resolution of the DNA bands:
a. Running the gel at a lower voltage for longer period of time
b. Using a wider gel comb
c. Loading less DNA into well
2. To get better separation of bands, adjust the agarose percentage of the gel if the similarly sized bands that bands that are running too close together are loaded. A higher percentage agarose gel will help resolving smaller bands from each other, and a lower percentage gel will help separating larger bands.
Protocol for 1% Agarose Gel
Staining Protocol
A. Pre-cast Protocol
1. Prepare agarose gel solution using your standard protocol.
2. Let the gel solution cool down and add the Gel RedSave fluorescent Stain10,000X stock into the agarose gel solution at 1:10,000 (for example, add 5 µl of dye to 50 ml of agarose solution) and mix thoroughly.
3. Cast the gel and allow it to solidify.
4. Load samples and perform electrophoresis using your standard protocol.
5. Detect the bands in the stained gel with a standard 300 nm UV transilluminator or blue LED illuminator.
Note: The pre-cast protocol is not recommended for polyacrylamide gels. Use the post staining protocol for acrylamide gels.
B. Post-staining Protocol
1. Run gels as usual according to your standard protocol.
2. Dilute the Gel RedSave fluorescent Stain10,000X stock reagent 5,000 fold to make a 2X staining solution in TBE or TAE buffer.
3. Carefully place the gel in a suitable polypropylene container. Gently add a sufficient amount of the 2X staining solution to submerge the gel.
4. Agitate the gel gently at room temperature for 30 min.
5. Image the stained gel with a standard UV transilluminator or blue LED illuminator.
Note: Optimal staining time and the amount of stain may depend on the thickness of the gel and the percentage of agarose.