| Product Description | Aladdin's Gel-Green dye can be used to stain DNA, RNA and other nucleic acids in gels, designed to replace the the highly toxic and mutagenic ethidium bromide (EB). It is superior to EB in noncytotoxicity, sensitivity, and stability. Nucleic acids stained by Gel-Green in gels are excited by blue light at about 500nm (UV light at about 260nm can also be used) to produce green fluorescence that can be examined by the gel imaging system used for SYBR Green or SYBR Gold detection.Gel-Green has a maximum excitation wavelength of about 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.Gel-Green is safer than EB or SYBR Green. Gel-Green is not cytotoxic or mutagenic at concentrations well above its working range. Gel-Green and Gel-Red, another safe nucleic acid dye, have special chemical structures that make then difficult to enter cells, thus greatly reducing or even avoiding the mutagenicity and 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.Gel-Green is highly sensitive and effective for staining nucleic acids with small molecular weight. When staining gel after electrophoresis, Gel-Green has similar or even higher sensitivity than SYBR Gold. Different from SYBR Gold, Gel-Green is also highly sensitive when precast in gels. The staining effect of Gel-Green of recommended concentration in this manual is slightly better than EB, and the working concentration of Gel-Green can be increased if higher sensitivity is desired.Gel-Green has good stability and high reproducibility of staining. The reproducibility of SYBR Green nucleic acid staining method is poor, usually due to its low stability. In contrast, Gel-Green is thermostable and has strong light resistance, and thus produces nucleic acid staining results with good reproducibility.Gel-Green can be examined with the same detection system as SYBR Green or EB. Gel-Green has almost the same optical properties as SYBR Green and SYBR Gold, with similar excitation and emission spectra, so Gel-Green can be used as a direct substitute for SYBR Green. The imaging system used for EB observation can be used for examining Gel-Red, but has lower sensitivity in detecting Gel-Green. For gel imaging systems assembled with both UV and blue light, we recommend replacing EB directly with Gel-Green. Please refer to Figure 1 for the excitation and emission spectra of Gel-Green.Figure 1. Excitation and emission spectra of Gel-Green .Gel-Green can be used in the same way as EB. Gel-Green 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 Gel-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. Gel-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 Gel-Green, dilute the stock to 1X with cell culture medium to stain cultured cells, followed by examination by fluorescence microscopy. High quality Gel-Green does not stain live cells, while the counterfeit does. Use blue light to examine Gel-Green stained gels. The way to distinguish between blue light and UV light is EB or Gel-Red stained nucleic acid is visible under UV light, but not under blue light. The prepared agarose gel containing Gel-Green can be stored at 4℃ in the dark for 3-5 days for future use.If the Gel-Green agarose gel will be used after melting, an appropriate amount of Gel-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 Gel-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.Gel-Green, like Gel-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 resolved 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.This product is compatible with commonly used electrophoresis buffers, such as TAE and TBE.Gel-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 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, Gel-Green can be used by the following two methods.1.Addition of Gel-Green in the gelPrepare agarose gel of the desired concentration (e.g., 1-3%). After the agarose is melted and properly cooled, add Gel-Green at a ratio of 1:10000 (e.g., add 10 µl of Gel-Green 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: Gel-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 Gel-Green staining solution by diluting Gel-Green with 100 mM NaCl solution or water at a ratio of 1:2500-1:5000 (e.g., add 20-40µl Gel-Green per 100 ml of water). After electrophoresis, immerse the agarose gel in an appropriate amount of Gel-Green staining solution, and stain gel for 20-30 minutes with slow shaking at 30-50rpm. 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 500nm or by other appropriate gel imaging systems. The Gel-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-Red1mlD0130NA-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
|
|---|
