99.5% Tris Saturated Phenol Formulated CAS.77-86-1 Color White

How is Tris saturated phenol formulated CAS.77-86-1 color white

Tris saturated phenol is a light yellow transparent liquid, the lower layer is the phenol phase, and the upper layer is Tris hydrochloric acid buffer pH>7.8. It is redistilled phenol saturated with Tris-HCl (pH 8.0). It has a pungent odor and contains Anti-oxidant 8-hydroxyquinoline. Tris saturated phenol is used to separate DNA.

The configuration of Tris saturated phenol: Take out the redistilled phenol from the refrigerator, place it at room temperature, and then melt in a 68°C water bath. Do not put it in a 68°C water bath immediately to prevent the glass from bursting. Add 8-hydroxyquinoline to 0.1% and β-mercapto Ethanol to 0.2%, (stock solution 14.4MOL/ML), mix well, the solution turns yellow, pour it into a separatory funnel (can also be carried out in a beaker), add 1MOL/ML Tris (PH8.0) of other solvents, repeat After mixing, let it stand still until it separates; release the lower yellow phenol liquid, discard the upper layer, add solid Tris about 1g/100ml, shake well, remove the water phase, add  0.1MOL/ML Tris (PH8.0) to balance several times, to pH 8.0, 0.1MOL/ML Tris (PH8.0) stored in a brown bottle at 4 degrees.

Tissue lysate, ensure that the pH is around 8.0 (add 0.1 volume of 1M Tris-HCl, pH 8.0, adjust it to around 8.0), and then add 0.5 volume of Tris saturated phenol (set aside during use, remove the lower phenol phase Use. The upper layer is a Tris-HCl solution to prevent phenol from contacting air and slow down the oxidation of phenol), 0.5 volume of chloroform: isoamyl alcohol (24:1), shake for 10 seconds and then ice bath for 15 minutes. After centrifugation, take the water phase (upper layer) and add an equal volume of isopropanol to precipitate the DNA, rinse it with 75% ethanol, and dissolve it in an appropriate amount of TE for later use.

Tris saturated phenol generally has a pH greater than 7.8 and is used for DNA extraction. Among them, phenol is a strong protein denaturant, which can denature and precipitate proteins in cells or tissues. The main function of Tris is to prevent and stop the oxidation of phenols. If phenols are oxidized, quinones (two benzene rings) will be formed. Contains strong free radicals, which can damage the structure of nucleic acids. At the same time, since the pH value is greater than 7, DNA is in the water phase and RNA is in the organic phase in an alkaline environment, so that the supernatant is separated and DNA can be obtained.

Tris Balanced Phenol is highly corrosive, so avoid skin contact or inhalation. If it turns red or brown, it indicates that oxidation has occurred and it cannot be used.

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