| Product Description | Aladdin's NA-Green Plus is an upgraded version of NA-Green, a new generation fluorescent nucleic acid dye designed to replace the highly toxic and mutagenic ethidium bromide (EB).NA-Green Plus is superior to EB in noncytotoxicity, sensitivity, and stability. The nucleic acid stained by NA-Green Plus exhibits green fluorescence when excited by blue light at approximately 500 nm, which can be examined by the gel imaging system originally used for the observance of SYBR Green or SYBR Gold. UV light at approximately 260 nm can also be used, but the nucleic acid band is relatively weaker.NA-Green Plus is an enhanced version of NA-Green. They are used in exactly the same way, but NA-Green Plus generates clearer and more even bands, lower background, and greatly reduced band tailing and bending. It does not affect nucleic acid migration, resulting in good band separation even with very high sample volumes.he gel containing NA-Green Plus is light orange. After electrophoresis, the color of the upper part of the gel is darker and the lower part is lighter, which is normal and has no effect on the electrophoresis results.NA-Green Plus has the maximum excitation wavelength of around 500 nm, which can be examined by human-friendly blue light, avoiding the mutagenicity of conventional UV on nucleic acid samples and the damage of UV to human eyes and skin.NA-Green Plus is safer than EB or SYBR Green. NA-Green Plus is not cytotoxic or mutagenic even at concentrations far above its working range. Its mutagenicity of the dye is far less than EB. NA-Green Plus, NA-Green and NA-Red, 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-Green Plus is highly sensitive and effective for staining nucleic acids with small molecular weight. NA-Green Plus can detect DNA samples down to 1 ng and is better 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-Green Plus has similar or even higher sensitivity than SYBR Gold. Different from SYBR Gold, NA-Green Plus is also highly sensitive when precast in a gel. The staining effect of NA-Green Plus of concentration recommended in this manual is slightly better than EB, and the working concentration of NA-Green Plus can be increased if higher sensitivity is desired.NA-Green Plus is stable and generates reproducible staining results. The reproducibility of SYBR Green nucleic acid staining method is poor, usually due to the low stability of the SYBR dyes. In contrast, NA-Green Plus is thermostable and has strong light resistance, and thus produces nucleic acid staining results with good reproducibility.NA-Green Plus can be examined with the same detection system as NA-Green, SYBR Green or EB. NA-Green Plus has almost the same optical properties as NA-Green, SYBR Green and SYBR Gold, with similar excitation and emission spectra, so NA-Green Plus can be used as a direct substitute for NA-Green or SYBR Green. The imaging system used for EB observation can be used for examining NA-Red, but has lower sensitivity in detecting NA-Green Plus. For gel imaging systems assembled with both UV and blue light, we recommend replacing EB directly with NA-Green Plus.Please refer to Figure 1 for the excitation and emission spectra of NA-Green Plus.Figure 1. The excitation and emission spectra of NA-Green Plus NA-Green Plus can be used in the same way as NA-Green and EB. Green Plus 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-Green Plus 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 Plus 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 Plus, 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-Green Plus does not stain live cells, while the counterfeit does.Use blue light to examine NA-Green Plus 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 Plus can be stored at 4ºC in the dark for 3-5 days for future use.If the NA-Green Plus agarose gel will be used after melting, an appropriate amount of NA-Green Plus 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 Plus 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-Green Plus, like NA-Green and NA-Red, 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 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 Plus is compatible with commonly used electrophoresis buffer solutions, such as TAE and TBE.NA-Green Plus 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 Plus can be used by the following two methods.1. Add NA-Green Plus to the gelPrepare agarose gel of the desired concentration (e.g., 1-3%). After the agarose is melted and properly cooled, add NA-Green Plus at a ratio of 1:10000 (e.g., add 10 µl of NA-Green Plus per 100 ml 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 Plus 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-Green Plus staining solution by diluting NA-Green Plus with 100 mM NaCl solution or water at a ratio of 1:2500-1:5000 (e.g., add 20-40µl NA-Green Plus per 100 ml of water). After electrophoresis, immerse the agarose gel in an appropriate amount of NA-Green Plus 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-Green Plus 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:产品编号产品名称包装D0071DNA Loading Buffer (6X)2mlD0072DNA Loading Buffer (Red, 6X)2mlD0128NA-Red 1mlD0130NA-Red5mlD0133NA-Green1mlD0135NA-Green5mlD0136-1mlNA-Green Plus1mlD0136-5mlNA-Green Plus5mlD0139Gel-Red0.2mlD0140Gel-Red1mlD0143Gel-Green0.2mlD0145Gel-Green1mlD0147-0.2mlGel-Green Plus0.2mlD0147-1mlGel-Green Plus1mlST004LAgarose50gST004MAgarose (Low EEO)50gST004QAgarose (Low EEO)250gST716TAE (50X)500mlST718TBE (5X)500mlST720TBE (1X premixed powder)2LST721TBE (1X premixed powder)10×2LST723TBE (5X premixed powder)2×2L
|
|---|
