| Product Description | Aladdin's NA-Green is an upgraded fluorescent dye to replace the highly toxic and mutagenic ethidium bromide (EB) for staining DNA and RNA in gels. NA-Green is superior to EB in noncytotoxicity, sensitivity, and stability. The nucleic acids stained by NA-Green exhibit green fluorescence when excited by blue light at approximately 500nm, which can be examined by a gel imaging system originally used for the observance of SYBR Green or SYBR Gold. UV light at approximately 260nm can also be used for the observation. NA-Green has a maximum excitation wavelength of around 500nm. The stained gels can be examined by blue light, avoiding the mutagenicity of conventional UV on nucleic acids and the damages of UV to human eyes and skin.NA-Green is safer than EB or SYBR Green. NA-Green is not cytotoxic or mutagenic even at concentrations well above its working range. NA-Green and NA-Red have special chemical structures that makes them difficult to enter cells, thus greatly reducing or even avoiding the mutagenicity and the cytotoxicity. 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-Green is highly sensitive and effective for staining nucleic acids with small molecular weight. The sensitivity of NA-Green in staining nucleic acids is 8-10 times of that of EB, especially when detecting small molecular weight DNA, making it better than EB for detecting trace amounts of DNA or RNA. When staining gel after electrophoresis, NA-Green has similar or even higher sensitivity than SYBR Gold. Different from SYBR Gold, NA-Green is also highly sensitive when precast in a gel. The staining effect of recommended NA-Green concentration in this manual is slightly better than EB, and the working concentration of NA-Green can be increased if higher sensitivity is desired.NA-Green is stable and generates reproducible staining results. The reproducibility of SYBR Green staining method is poor, usually due to the its low stability. In contrast, NA-Green is thermostable and has strong light resistance, and thus produces nucleic acid staining results with good reproducibility.NA-Green can be examined with the same detection system as NA-Green, SYBR Green or EB. NA-Green has almost the same optical properties as SYBR Green and SYBR Gold, with similar excitation and emission spectra, so NA-Green can be used as a direct substitute for SYBR Green. The imaging system used for EB observation can be used for examining NA-Red, but has lower sensitivity in detecting NA-Green. For gel imaging systems assembled with both UV and blue light, we recommend replacing EB directly with NA-Green. Please refer to Figure 1 for the excitation and emission spectra of NA-Green.Figure 1. The excitation and emission spectra of NA-Green NA-Green can be used in the same way as EB. NA-Green can be added directly to agarose gels 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-Green 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. NA-Green 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-Green, dilute the stock to 1X with cell culture medium to stain cultured cells, followed by examination by fluorescence microscopy. High quality NA-Green does not stain live cells, while the counterfeit does. Use blue light to examine NA-Green stained nucleic acid gels. The way to distinguish between blue light and UV light is EB or NA-Red stained nucleic acid is visible under UV light, but not under blue light. The prepared agarose gel containing NA-Green can be stored at 4℃ in the dark for 3-5 days for future use.If the NA-Green agarose gel will be used after melting, an appropriate amount of NA-Green 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-Green 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.Both NA-Green and NA-Red can stain double-stranded DNA (dsDNA) as well single-stranded DNA (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 resolved well, we recommend staining gel 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-Green is compatible with commonly used electrophoresis buffers, such as TAE and TBE.NA-Green is suitable for imaging systems with excitation light of about 500nm or 260nm. The fluorescence can also be observed well with ordinary UV lamps or imaging systems suitable for NA-Red or EB (with the excitation light of about 300nm), but the intensity will be slightly weakerNA-Green 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-Green can be used by the following two methods.1.Addition of NA-Green to the gelPrepare agarose gel of the desired concentration (e.g., 1-3%). After the agarose is melted and properly cooled, add NA-Green at a ratio of 1:2000 (e.g., add 50µl of NA-Green 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 blue light at approximately 500nm. Note: NA-Green 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.Staining gel after electrophoresisPrepare the NA-Green staining solution by diluting NA-Green with 100mM NaCl solution or water at a ratio of 1:500-1:1000 (e.g., add 100-200µl of NA-Green per 100ml of water). After electrophoresis, immerse the agarose gel in an appropriate amount of NA-Green 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 under 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 under blue light with excitation wavelength of approximately 500nm or by other appropriate gel imaging systems. The NA-Green 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.Related Products:Cat. No.Product NamePack SizeD0071DNA Loading Buffer (6X)2mlD0072Red DNA Loading Buffer (6X)2mlD0128NA-Red (2000X)1mlD0130NA-Red (2000X)5mlD0133NA-Green (2000X)1mlD0135NA-Green (2000X)5mlD0136-1mlNA-Green Plus (2000X)1mlD0136-5mlNA-Green Plus (2000X)5mlD0139Gel-Red (10000X)0.2mlD0140Gel-Red (10000X)1mlD0143Gel-Green (10000X)0.2mlD0145Gel-Green (10000X)1mlD0147-0.2mlGel-Green Plus (10000X)0.2mlD0147-1mlGel-Green Plus (10000X)1mlST004LAgarose50gST004MAgarose (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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