What is the role of tris saturated phenol

Tris saturated phenols are generally pH greater than and are used for DNA extraction. Among them, phenols are strong protein denaturants that can denature proteins in cells or tissues.

Precipitation. The main function of TRIS is to prevent and control phenolic oxidation, if phenolic oxidation, it will form quinone (two benzene rings, quinone contains strong free radicals.

It will destroy the structure of nucleic acids.

At the same time, since the pH value is greater than 7, in an alkaline environment the DNA is in the aqueous phase and the RNA is in the organic phase. The supernatant is thus separated, and the DNA is obtained.

Configuration of Tris saturated phenol:

1. Take out the re-steamed phenol from the refrigerator and dissolve it in a 68-degree water bath at room temperature, and do not put it in a 68-degree water bath immediately to prevent the glass from bursting.

2. Add 8 hydroxyquinoline to mercaptoethanol.

to, (stock solution 14.)4mol ml), mix well, the solution is yellow, pour into a separating funnel (also in a beaker.

3. Add 1mol ml of tris ( of the same solvent, mix it repeatedly, and then stand to stratify; Release the lower layer of yellow phenol solution and discard the upper layer.

4. Add about 1g of 100ml phenol to the solid tris, shake well, and remove the aqueous phase.

5. Add balance several times to pH.

6. Add to the brown bottle and store at 4 degrees.

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

Hello, glad to answer for you!

The main function of TRIS is to prevent and control phenolic oxidation, if phenolic oxidation, it will form quinone (two benzene rings), which contains strong free radicals and will destroy the nucleic acid structure.

Tris saturated phenols are generally pH greater than and are used for DNA extraction. Among them, phenol is a strong protein denaturant that can denature and precipitate proteins in cells or tissues.

Hope you help!

Tris-HCl is a buffer system, and Tris-saturated phenol can ensure that the phenol solution can play a role in a stable pH environment. And water does not have the ability to buffer.

Tris-HCl saturated phenol has a certain acid-base buffering effect.

1. Take out the resteamed phenol and dissolve it in a 68-degree water bath at room temperature, and do not put it into a 68-degree water bath immediately to prevent the glass from bursting.

2. Add 8 hydroxyquinoline to and mercaptoethanol to the stock solution mol ml), mix the mountain and open evenly, the solution is yellow, and pour it into the separating funnel (can also be carried out in a beaker).

3. Add 1 mol ml of Huweidong Tris() with equal solvent, mix it repeatedly, and then stand to stratify; Release the lower layer of yellow phenol solution and discard the upper layer.

4. Add about 1g of 100ml phenol to the solid tris, shake well, and remove the aqueous phase.

5. Add mol mltris (pH balance several times, until pH is 6, add mol mltris (pH is stored at 4 degrees in a brown bottle.