Hello
Does anyone have a method for the preparation of the solution of phenol saturated with Tris-HCL pH = 7.5? I thank the person who can help me.
Hi there,
Handling of crystallized phenol is hazardous then buying the ready to use tris saturated phenol is the most sensible move to do. Otherwise, melt the phenol at 65°C and then add 1 volume of 10x concentrated buffer and mix thoroughly. Wait until phase separation and decant. Renew this several times with 1x buffer until the pH of the aqueous phase is stable. Then store the phenol solution covered by a thin layer of buffer at 4°C.
Melt phenol, add equal volume 100mM Tris pH 7.5, mix, allow aqueous phase to come to the top (you can spin these in 50 ml tubes), remove top and add fresh 100mM Tris pH 7.5. Repeat three times. After removing the last Tris treatment from the top, wash the phenol twice with water. Removing excess Tris will improve the shelf life. Store phenol under water in a light-proof container in the cold (4C); remember Phenol is on the bottom. Make sure the container is sealed well.
And do all this under a fume hood with proper gloves (nitrile, not latex)
I agree completely with Dominique. And Catherine: read and follow ALL of the safety advice: phenol is awful stuff to work with! And remember that it dissolves polystyrene, including the lids of 50mL Sterilin (etc) centrifuge tubes!!
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