Is Too Much Phosphatase Bad?

Ian A. York iayork at
Tue May 13 07:56:48 EST 1997

Robert A. Schoenfeld <schoenfeld at MSSCC.MED.UTAH.EDU> wrote:
>Are there any deleterious effects when too much phosphatase is used - 
>i.e. ragged ends?

I don't know exactly what the damage is, but from experience
overphosphatasing will drasticly reduce ligation efficiency.  I used to
avoid phosphatasing at all costs, until (d'oh!) I went through the
calculations and realized that I was over-treating by a factor of 100 or
so because I was just using 1 ul; I had just assumed that, as with most
enzymes, a moderate excess was no big deal. When I started calculating the
correct amount, which meant diluting the phosphatase way down, the
problems disappeared. 


      Ian York   (iayork at  <>
      "-but as he was a York, I am rather inclined to suppose him a
       very respectable Man." -Jane Austen, The History of England

More information about the Methods mailing list