Why use tris saturated phenol instead of phenol directly

The main reasons for using tris-saturated phenol instead of directly using phenol are as follows:

Phenol is easily oxidized in the air, and the quinones formed after oxidation will bind firmly to the DNA strand, which will cause the extracted DNA to be unable to be used for subsequent operations, such as PCR or enzyme digestion.

Phenol oxidation of quinones and other products can cause cleavage of phosphodiester bonds and cross-linking of RNA and DNA, and direct use of unsaturated phenol may result in a decrease in the quality of the extracted DNA.

Tris-saturated phenols are better at maintaining pH stability. Because pH is critical for nucleic acid extraction, different pH values can affect the solubility and stability of nucleic acids.

Tris-saturated phenols provide better protection of nucleic acids. Because Tris-saturated phenol can effectively resist the influence of external acids and alkalis, maintain the stability of the solution, and thus better protect nucleic acids.

Therefore, in order to improve the quality and efficiency of nucleic acid extraction, tris-saturated phenol is often used instead of phenol directly.

Commercially available phenols contain oxides such as quinones, which can cause phosphodiester bonds.

The breakage and resulting cross-linking of RNA and DNA should be re-evaporated with a condenser tube at 160 °C. Re-distilled phenol was added with 8-hydroxyquinoline (as an antioxidant) and repeatedly extracted with equal volumes of Tris·Cl ( and Tris·Cl ( buffer to saturate and make it pH.

Reach the above, for tris saturated phenol (left and right).

There are two kinds of saturated phenols in nucleic acid purification, one is Tris saturated phenol (left and right), so it is also called basic phenol. The other is acidic phenols, which are water-saturated phenols. For RNA extraction, acidic phenols, that is, water-saturated phenols, are prepared, because under acidic conditions, RNA is assigned to the aqueous phase, while DNA is in the organic phase, so as to achieve the purpose of isolating RNA.

Water-saturated phenols can also denature proteins.

precipitation, which separates the protein from the nucleic acid extract. Therefore, different pH values and different applications must not be confused to avoid experimental failure.

1. Take out the solid phenol and dissolve it in a 68-degree water bath at room temperature, and do not immediately put it into a 68-degree water bath to prevent the glass from bursting. 2. Add 8 hydroxyquinoline to and mercaptoethanol to the stock solution, mix well, the solution is yellow, and pour it into the separating funnel (it can also be carried out in a beaker). 3. Add 1mol ml of tri of the same solvent

Phenol can denature proteins, have a strong corrosive effect on mucous membranes, and can also inhibit the central nervous system or damage liver and kidney function.

Burns on **: The wound is initially white wrinkled, and then a brown crust is formed.

A small amount of phenol splashed on the ** will not cause particularly serious injury, just pay attention to it in the future Be sure to wear gloves when operating in the future, if you accidentally drip it, quickly rinse it with a large amount of flowing water for at least 20 minutes; If the area is small, the wound can also be wiped with 50% alcohol or wiped with glycerin, polyethylene glycol or a mixture of polyethylene glycol and alcohol (7:3)**, and then rinsed with plenty of running water immediately. Then apply a wet compress with saturated sodium sulfate solution.

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!

In this way, phenol is very susceptible to oxidation in the air, and the quinones formed after oxidation will bind firmly to the DNA strand, and in the most serious case, the extracted DNA will not be able to perform subsequent operations (PCR or enzyme digestion). It is recommended that if you must use it, re-steam the phenol and saturate it with water or Tris hydrochloric acid. However, this step is too complicated and is generally not recommended to use it, because Tris saturated phenol is really cheap.

If it is urgent, I personally recommend that you do not use phenol at all, and use chloroform isoamyl alcohol directly. The CTAB method is generally used for the extraction of plant genomic DNA, and the absence of phenol extraction generally does not have much effect.