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CelGreen nucleic acid Dye
CAT：CG**1
U.S.＄*0.*0 for **0ul （negotiable） Storage：Room temperature and away from light.  
Characteristics of CelGreen nucleic acid dye
● Non-toxic: CelGreen's unique oiliness and large molecular weight make it unable to penetrate the cell membrane into the cell, and Ames' test results also show that the mutagenicity of the dye is far less than EB.
● High sensitivity: suitable for electrophoretic staining of fragments of various sizes, with less effect on nucleic acid migration than SYBR Green I.
● High stability: suitable for using microwave or other heating methods to prepare agarose gel; Extremely stable in acid or alkali buffers at room temperature, strong light resistance.
● High signal-to-noise ratio: sample fluorescence signal is strong, background signal is low.
● Simple operation: Like EB, the dye does not degrade in the process of preforming glue and electrophoresis; The dyeing process after electrophoresis also takes only *0 minutes without decolorization or rinsing, and can be directly observed with the ultraviolet gel transmisometer.
● Wide range of application: you can choose pre-electrophoresis dyeing (glue dyeing) or post-electrophoresis dyeing (bubble dyeing); Suitable for agarose gel or polyacrylamide gel electrophoresis; Can be used for dsDNA, ssDNA or RNA staining.
● The best excitation can be obtained near **4nm visible light.
Usage
1. Glue dyeing method (same as EB) (recommended method)
(1) Add CelGreen nucleic acid dye when making glue (for example: add 5μL CelGreen *0,**0× storage solution per *0mL agarose solution, and so on).
(2) Electrophoresis was performed according to conventional methods.
Note:
The amount of dye used in this method is relatively small. **0 μL of dye can make about **0 pieces of *0mL glue.
Because CelGreen has good thermal stability, it can be added directly in hot agarose solution without waiting for the solution to cool. Shake, oscillate, or flip to ensure that the dye is well mixed. Alternatively, the CelGreen reservoir can be added to the agarose powder and electrophoresis buffer, and then heated in the microwave or other common method to prepare the agarose gel. CelGreen is compatible with all commonly used electrophoresis buffer solutions.
If you always see band dispersion or separation is not ideal, it is recommended to use bubble dyeing to confirm whether the problem is related to the dye. If the problem persists after dyeing, the problem is not related to the dye, please try: reduce the agarose concentration; Use longer gels; Extend the gel time to ensure clear edges; Improve the sampling technique or choose blister dyeing method.
This method is not suitable for prefabricated polyacrylamide gel, please use the foam dyeing method for polyacrylamide gel.
 
2. Blister dyeing
(1) Electrophoresis was performed according to conventional methods.
(2) The CelGreen *0,**0× storage solution was diluted about 3,**0 times with H2O into 0.1M NaCl to make a 3× dyeing solution. (For example, *5μL CelGreen *0,**0× reservoir and 5mL 1M NaCl are added to *5mL H2O).
(3) Place the gel carefully into a suitable container, such as a polypropylene container. Slowly add a sufficient amount of 3× dye solution to immerse the gel. The optimal dyeing time varies slightly according to the gel thickness and agarose concentration. For gels containing 3.***0% acrylamide, the dyeing time is usually between *0min and 1h, and increases with the increase of acrylamide content.
(4) The results were observed by **0nm visible light system.
Note:
When dyeing with bubble dyeing method, the amount of dye is more. A single use of the dye solution can be reused for about 3 times.
3× CelGreen dyeing solution can be prepared in large quantities and stored away from light at room temperature until used up.
3. PAGE steps for nucleic acid electrophoresis:
(1) Put the gel prepared by TBE into the electrophoresis tank and hold the edge with a clamp.
(2) Fill the buffer tank with 5×TBE in the same batch as the gel solution. Use a syringe to remove air bubbles from the bottom of the gel.
(3) Use a syringe to draw 1×TBE to flush the sample addition hole. The DNA sample was mixed with an appropriate amount of 6× gel loading buffer, and added into the sampling hole with a micro-pipette.
(4) Connect the electrode to the power supply (positive terminal slot), turn on the power supply generally *0V; 1 to 8V/cm. Electrophoresis was performed for 9h.
(5) Electrophoresis until the standard reference dye migrates to the desired position (generally, electrophoresis until the xylene is completely removed, and the bromophenol blue is stopped 2 ~ *5px from the bottom edge). Turn off the power, unplug and discard the electrophoresis solution from the bath.
(6) Take the gel off and put it in the dyeing dish, add 3XCelGreen 1X buffer for oscillating dyeing for ****0 minutes, and place it in the ultraviolet detection.
Note:
PAGE glue is different from aglyceride sugar gel, which can not be pre-dyed or spot-dyed. Can only use foam dyeing method color, because polyacrylamide is relatively dense, the dye is not easy to go deep, the color effect is not as good as Joan ester sugar gel.
Special reminder:
1. If you are using an ultraviolet imager, please select CelRed; If you use a laser imager or wish to observe in visible light, choose CelGreen.
2. In rare cases, the DNA sample after the plasmid is cut by certain enzymes will have a tailing and resolution reduction, in which case it is recommended to try two staining methods at the same time to determine which method is more appropriate.