| Product Description | Aladdin's NA-Red is a new generation fluorescent nucleic acid dye designed to replace the highly toxic and mutagenic ethidium bromide (EB).NA-Red is superior to EB in noncytotoxicity, sensitivity, and stability. The nucleic acid stained by NA-Red exhibits red fluorescence when excited by UV light at approximately 300nm, which can be examined by the gel imaging system originally used for the observance of EB.NA-Red is safer than EB or SYBR Green. NA-Red is not cytotoxic or mutagenic even at concentrations far above its working range. The Ames test also proves that the mutagenicity of NA-Red is far less than that of EB. NA-Red and NA-Green have a special chemical structure that makes it difficult to enter cells, thus greatly reducing or even avoiding the mutagenicity and cytotoxicity of the dye. SYBR Green, on the other hand, can penetrate cell membranes, enter live cells and stain DNA, and has been reported to strongly enhance UV-induced mutagenesis.NA-Red is highly sensitive and effective for staining nucleic acids with small molecular weight. NA-Red is 8-10 times more sensitive than EB for detecting trace amounts of DNA or RNA, and is especially sensitive for small molecular weight DNA. When staining gel after electrophoresis, NA-Red has similar or even higher sensitivity than SYBR Gold. Different from SYBR Gold, NA-Red is also highly sensitive when precast in a gel. The staining effect of NA-Red of concentration recommended in this manual is slightly better than EB, and the working concentration of NA-Red can be increased if higher sensitivity is desired.NA-Red has good stability and high reproducibility of staining. The reproducibility of SYBR Green nucleic acid staining method is poor, usually due to the low stability of the SYBR dyes. In contrast, NA-Red is thermostable and has strong light resistance, and thus produces nucleic acid staining results with good reproducibility.NA-Red can be examined with the same detection system as EB. NA-Red has almost the same excitation and emission spectra as EB, so it can be used as a direct substitute for EB. Please refer to Figure 1 for the excitation and emission spectra of NA-Red.Figure 1. The excitation and emission spectra of NA-Red NA-Red can be used in the same way as EB. NA-Red can be added directly to agarose gel after melting in an appropriate proportion, or the gel can be stained after electrophoresis is completed. The former method is more convenient, while the latter is a bit more sensitive. However, since NA-Red itself is already very sensitive, it is usually sufficient to precast it in the gel. For some special cases, such as samples with particularly small amounts of nucleic acid, gel staining after electrophoresis is recommended.The effect of NA-Red on the mobility of nucleic acids is very small and less than that of SYBR Green.NA-Red bound to DNA or RNA can be effectively removed by gel recovery kits or phenol/chloroform extraction, thus enabling the subsequent ligation, digestion, PCR, sequencing, and other routine molecular biology applications.
Precautions： To check the quality of NA-Red, dilute the stock to 1X with cell culture medium which is then used to stain cultured cells, followed by examination by fluorescence microscopy. High quality NA-Red does not stain live cells, while the counterfeit does.The prepared agarose gel containing NA-Red can be stored at 4ºC in the dark for 3-5 days for future use.If the NA-Red agarose gel will be used after melting, an appropriate amount of NA-Red needs to be added for better observation.Reuse of the gel after sample loading and electrophoresis is not recommended.Gels that are stained with NA-Red after electrophoresis generally do not need to be decolorized prior to examination. If the background is too high, decolorization can be performed using nuclease-free water.NA-Red, like NA-Green, can stain dsDNA, ssDNA and RNA.For polyacrylamide gels, stain after electrophoresis and extend the staining time to 30 minutes - 1 hour.If DNA smear is observed or DNA could not be separated well, we recommend gel staining after electrophoresis to determine whether the problem is due to the dye. If the problem still exists other attempts can be tried, such as using freshly prepared buffer, reducing the amount of nucleic acid loaded, reducing the concentration of dye or agarose, using a longer gel, reducing the voltage for electrophoresis and extending the electrophoresis time to improve electrophoresis.NA-Red is compatible with commonly used electrophoresis buffer solutions, such as TAE and TBE.NA-Red is not a toxic or hazardous substance and has passed environmental safety related tests. The related waste does not require special treatment and can be disposed of as conventional chemical reagents.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.
Instructions for Use： Similar to EB, NA-Red can be used by the following two methods. 1. Add NA-Red to the gel Prepare agarose gel of the desired concentration (e.g., 1-3%). After the agarose is melted and properly cooled, add NA-Red at a ratio of 1:10000 (e.g., add 10µl of NA-Red per 100ml of gel). Mix well, and pour the gel. After electrophoresis of DNA or RNA samples, bright nucleic acid bands in the gel should be visible when excited by UV light.Note: NA-Red has good thermal stability, so it can be added directly to the hot agarose solution without cooling. It also can be mixed together with the agarose powder and the electrophoresis buffer prior to melting.2. Stain gel after electrophoresisPrepare the NA-Red staining solution by diluting NA-Red with 100 mM NaCl solution or water at a ratio of 1:2500-1:5000 (e.g., add 20-40µl NA-Red per 100 ml of water). After electrophoresis, immerse the agarose gel in an appropriate amount of NA-Red staining solution, and stain gel for 20-30 minutes with slow shaking (30-50rpm) on a shaker. The staining time depends on the thickness of the gel. The thicker the gels, the longer the staining time. After staining, the nucleic acid bands can be examined by blue light. To obtain clearer bands, the stained gel can be rinsed 1-2 times with water for 3-5 minutes each time to eliminate the background and then examined by blue light with excitation wavelength at approximately 500 nm or by other appropriate gel imaging systems. The NA-Red staining solution can be reused about 3 times or prepared in large quantities at one time and stored at room temperature in the dark. For cases where nucleic acids need to be recovered, care needs to be taken during operation to avoid nuclease contamination.Appendix: How to Quickly Distinguish EB (Ethidium Bromide) from NA-RedSince EB and NA-Red solutions are similar in color and are examined by the same detection system after nucleic acid staining, there is a particular need for a simple method to quickly differentiate between EB and NA-Red, besides the liquid chromatography-mass spectrometry (LC-MS) approach determining their molecular weight. We found that EB and NA-Red can be distinguished quickly by measuring the fluorescence intensity at a fixed emission wavelength when scanned within a range of excitation wavelength. As shown in Figure 2, the emission wavelength was fixed at 600 nm, and the excitation wavelength was ranged from 240 to 360 nm. It was found that different concentrations of free EB had an excitation maximum at 300 nm, and the overall scanning spectra were very clear (Figure 2A), which was basically consistent with the plasmid DNA-bound EB (Figure 2B). However, free NA-Red was hardly excited (Figure 2A), while the plasmid DNA-bound NA-Red had the maximum fluorescence intensity when excited at 300nm (Figure 2B). Therefore, EB and NA-Red in their free state can be differentiated by fluorescence examination.Figure 2. The fluorescence intensity of EB, NA-Red and their respective bound forms with plasmid DNA, when excited in the range of 240-360nm. A. The fluorescence intensity of free EB and NA-red. B. The florescence intensity of EB and NA-red binding to plasmid DNA. Related Products:产品编号产品名称包装D0071DNA Loading Buffer (6X)2mlD0072DNA Loading Buffer (Red, 6X)2mlD0128NA-Red 1mlD0130NA-Red5mlD0133NA-Green1mlD0135NA-Green5mlD0139Gel-Red0.2mlD0140Gel-Red1mlD0143Gel-Green0.2mlD0145Gel-Green1mlST004LAgarose50gST004MAgarose (Low EEO)50gST004QAgarose (Low EEO)250gST716TAE (50X)500mlST718TBE (5X)500mlST720TBE (1X premixed powder)2LST721TBE (1X premixed powder)10×2LST723TBE (5X premixed powder)2×2L
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