99% Min White Tris(Hydroxymethyl)Aminomethane CAS No 77-86-1

The difference between trimethylol aminomethane saturated phenol and water saturated phenol

Tris (trihydroxymethyl aminomethane) as a commonly used biological buffer has a very wide range of applications in the field of biochemical industry, such as nucleic acid purification. There are two kinds of saturated phenols in nucleic acid purification. One is Tris saturated phenols, also known as basic phenols, Tris balanced phenols (tris saturatedphenol or Tris balanced phenols), which are light yellow transparent liquid saturated with tris clph8.0 The upper layer was Tris hydrochloric acid buffer, and the antioxidant 8-hydroxyquinoline was added into the solution. It is suitable for removing protein and extracting DNA from nucleic acid samples.

The pH of Tris saturated phenols is generally higher than 7.8, among which phenol is a strong protein denaturant, which can denaturate and precipitate proteins in cells or tissues. The main function of Tris is to prevent and control the oxidation of phenols. If phenols are oxidized, quinones (two benzene rings) will be formed. Quinones contain strong free radicals, which will destroy the structure of nucleic acids. At the same time, due to the pH value greater than 7, DNA is in the aqueous phase and RNA is in the organic phase in alkaline environment. The supernatant was separated and DNA was obtained.

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The other is acidic phenol, or water saturated phenol, which is used to extract RNA. It is also called water saturated phenol or water balanced phenol. The pH of water saturated phenol is generally 4.7-5.5. It is usually used together with guanidine isothioate (or 0.1%, 0.2% β - mercapto, ethanol) to extract RNA from cells, so that DNA is in phenol phase and RNA is in water phase. The two are separated.

Configuration of Tris saturated phenol:

1. Take out the re steamed phenol from the refrigerator and place it in the 68 ℃ water bath at room temperature to dissolve it. Do not put it in the 68 ℃ water bath immediately to prevent the glass from cracking;

2. Add 8-hydroxyquinoline to 0.1% and β - mercaptoethanol to 0.2% (stock solution 14.4mol/ml), mix well, the solution turns yellow, and pour it into the separating funnel (it can also be carried out in a beaker);

3. Add 1mol / ml tris (ph8.0) of the same solvent, mix it repeatedly, and let it stand still until it is layered; release the Yellow phenol solution in the lower layer and discard the upper layer;

4. Add solid Tris about 1g / 100ml phenol, shake well and remove the aqueous phase;

5. Add 0.1mol/mltris (ph8.0) to balance several times until pH is 8.0;

6. Add 0.1mol/mltris (ph8.0) to brown bottle and store at 4 ℃;

7. If the Yellow disappears or turns pink (indicating that the phenol has been oxidized), it cannot be used.

The re evaporated phenol is a white needle like crystal with strong corrosiveness. It should be avoided to contact with skin or inhale into the body as far as possible. If it turns red or brown, it indicates oxidation and cannot be used. The storage condition should be kept away from light at - 20 ℃. Tris saturated phenols are toxic and irritating substances, which need to be operated in the fume hood.