Selection of nucleic acid gel electrophoresis dyes.

gel electrophoresis is our most basic molecular experiment. The basic principle is that the electrophoresis molecule moves under the action of the electric field, so it is in positive ratio to the strength of the electric field and the net charge of the electrophoresis molecule itself. Because of the use of non-reactive stable support medium in electrophoresis, which reduces converse motion, the migration rate of electrophoresis is inversely inversely compared with the coefficient of friction of molecules. The most commonly used
agar
sugar gel electrophoresis dyes in the lab today are EB, GeneFinderTM, GoldViewTM, Sybr Green I, GelRed, and Gel Green. The following analysis
of these
nucleic acids and dyes.
1.EB
Acetylene Ingot (EB) has become the most commonly used nucleic acid dye due to its inexpensive and easy-to-use, and is widely used to observe and detect
DNA
or RNA in agarose gels and polyacrylamide gels. However, EB is a strong mutagent, with moderate toxicity, irritating effects on the skin, eyes, mouth and upper respiratory tract system, potentially harmful to the human body. And the waste EB dye is easy to pollute the environment.
EB itself does not glow under ultraviolet light, and when combined with single-stranded, double-stranded, or triple-stranded DNA, it emits bright fluorescence, and EB-DNA bindings can cause light bleaching of dyes and DNA single-stranded fractures, with potentially mutagel effects. EB is simple to use, it can be added to the pre-dyeing when making glue, or after electrophoresis is completed. Front dyeing is conducive to saving time, but easy to appear strip deformation tail and other problems, after dyeing more time-saving, but the effect is better.
2.GeneFinder
. GeneFinderTM is a new type of nucleic acid dye that can replace EB as a dye for various nucleic acid electrophoresis. Unlike the highly carcinogenic EB, GeneFinderTM is an antholic dye with low toxicity, and GeneFinderTM combines with dsDNA to emit fluorescence, which detects nucleic acids with a higher sensitivity than EB staining.
GeneFinderTM dye has a maximum absorption peak of 470nm, the nucleic acid combined with the dye is green fluorescent under ultraviolet light, in addition, the dye can also cause
organic
body
mutation
, when used to take appropriate protective measures.
3.GoldView
. GoldViewTM is also a new nucleic acid dye that replaces ethyl bromide (EB), which, when dna is detected using agar glycositose electrophoresis, produces a strong fluorescent signal when combined with nucleic acids, which is as sensitive as EB and is used in exactly the same way. GoldView is good for staining large fragments of DNA, but not ideal for fragments below 500 bp, and its fluorophoric substrings are very easy to quench under UV lamps, generally about 5-10min bands will disappear. Therefore, as soon as possible to take pictures, observation. In addition, GoldView has poor sensitivity and a severe background color, which is not suitable for adhesive recycling. And its toxicity is greater, especially under ultraviolet lamps, its ability to induce mutation is very high.
4.Sybr Green I
. Sybr GreenI is also a commonly used nucleic acid dye that, when combined with double-stranded DNA (dsDNA), is greatly enhanced in fluorescence and is more sensitive than EB. Sybr GreenI, which is joined with double-stranded DNA, presents bright green fluorescence in ultraviolet perspective, but its poor stability (fear of light, fear of heat) makes dyeing less repetitive. It is also less sensitive to detecting DNA fragments less than l00 bp due to the lack of staining ability of DNA fragments below 50 bp. Sybr GreenI is just as simple to use and less denatured than EB, but it still doesn't guarantee the safety of the experimenter, and its high price limits its widespread use.
5. GelRed and GelGreen
GelRed and Gel Green are excellent fluorescent nucleic acid dyes that combine high sensitivity, safety and stability. It is safely determined by the U.S. Environmental Protection Agency that waste can be dumped directly into sewers without causing any environmental pollution. Most commercial nucleic acid dyes today are not entirely satisfactory in terms of sensitivity, stability and safety, and the listing of GelRed and Gel Green has changed that. Both GelRed and Gel Green show excellent experimental results, both for pre-made gel dyeing and post-gel electrophoresis dyeing.
GelRed
designed to work with
UV gel imaging systems, is as easy to use as EB. GelRed is still extremely sensitive when used in prefabribribrio gels and is also highly sensitive in detecting low concentrations and trace amounts of DNA, especially for small molecular weights.
Gel Green can be used by researchers using a blue visible laser gel scanner or a visible-light-excited DarkReader. GelGreen is superior to Sybr GreenI in both prefabribrio and post-gel electrophoresis dyeing, and there is no instability in the latter. Both GelRed and GelGreen have strong stability, can be stored at room temperature for a long time, and the thermal stability of these two dyes is also very high, in the electrophoresis buffer solution, can be used in the microwave oven
heating
without denaturation. Prefabribrilated gels containing dyes can be prepared in batches and stored for long periods of time.
GelRed and Gel Green are significantly more secure than other nucleic acid dyes such as EB or Sybr GreenI. Standard Ames test results conducted by independent testing services show that the special chemical structure of GelRed and Gel Green makes it difficult to penetrate the cell membrane into the cell, which ensures the safety of the experimenter and gives the experimenter more confidence to do the experiment.
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