Protocol Online logo
Top : Forum Archives: : Molecular Biology

70% Ethanol... - (Aug/22/2006 )

I did a Phenol/chloroform extraction today. But unfortunatly, I added 70% ethanol instead of 100% ethanol when doing ethanol precipitation. What's the worst consequence can be, and what can I do to make it up? sad.gif
Thx...

-josophen-

Worst: you'll never recover your DNA.

If you have space left add 100% EtOH until you get a final conc of 70%. If not then transfer to a bigger tube add 100% EtOH to 70% final volume and then split the mixture into smaller tubes to spin down.

-Doc_Martin-

And will it still lower the yield to a great extent in this way?
Do you mean the conc of Ethanol? Then how about the conc of NaAc?...
I really worry about the DNA now...

-josophen-

If you can somehow get the final volume of EtOH in your sample up to 70% (by adding 100% EtOH) you should be able to retrieve most of your DNA.

As for NaOAc, add 1/10th volume of 3 M.

-Doc_Martin-

Thanks a lot!! smile.gif Hope I won't be punished too hard.

-josophen-