Simply place the bottle of crystalline phenol in a 50c water bath, it is not necessary to melt all the penol. Transfer melted phenol to an appropriate bottle, then add 8-hydroxylquenaline to a final concentration of 0.1% (W/V).

Add equal volume of 200mM (or 250 mM) Tris base solution (it makes a seperated phase).Mix on magnetic stirrer, over night. Check the pH by paper pHmeter. it should be 8, otherwise repeat the procedure with 100mM Tris. Tris phase should remain on the phenol.

"....Crystalline phenol is not recommended because it must be redistilled at 160°C to remove oxidation products,..."

Sambrook et al., Molecular Cloning, A Laboratory Manual !!!

crysatalline phenol will not give the correct result, you should use water saturated or triss saturated eqilibrated phenol

Simply place the bottle of crystalline phenol in a 50c water bath, it is not necessary to melt all the penol. Transfer melted phenol to an appropriate bottle, then add 8-hydroxylquenaline to a final concentration of 0.1% (W/V).

Add equal volume of 200mM (or 250 mM) Tris base solution (it makes a seperated phase).Mix on magnetic stirrer, over night. Check the pH by paper pHmeter. it should be 8, otherwise repeat the procedure with 100mM Tris. Tris phase should remain on the phenol.

and always wear gloves while handling phenol..agree with Mohsen Basiri on the procedure of making tris saturated phenol

Why don't you ask your boss phenol already ready to use? it is safe for you and for all people around you in your Institute. Buffered phenol is not so expensive now. I did several time that work ....but 20 years ago...

If you are running low on bidget; then I will recommend using tris base for the equilibration of phenol.

You need to first melt the crtalline phenol using water bath, then wash it with progressive decreasing molar solution of trsi base (Ph 8) starting with 0.5M then 0.2 M and then two washings with 0.1M till it attains PH of 8 itself which you can use for DNA separation.

You can however use direct DNA extraction kits which ocst not more than 8000INR.

Regards

You can visit my profile and download one of my papers on DNA extraction

While saturating, if the pH goes above 8 then will it make any difference?

I have been doing this but facing difficult to separate the phase.  

Now-a-days Phenol:Chloroform:isoamylalcohol (25:24:1) together is commercially available .

We need to add phenol as an inducer to 1mM final concentration to some petri dishes, but I can't remember how to do it. Any help?

What of preparing just 40% phenol ? I weighed 40 grams of phenol and added 100 ml of distilled water but it did not dissolve but when I applied some heat it dissolved.However,it returned to the more state.Can anybody please suggest a better way of doing it.I actually want to extract with 40% hot phenol (70 degrees Celsius).Thanks.