PCR cloning - PAGE

Julian Parkhill J.Parkhill at bham.ac.uk
Wed Aug 23 09:10:15 EST 1995

In article <1995Aug22.215624.59506 at cc.usu.edu>, jmorrey at cc.usu.edu (J.
Morrey) wrote:

> Will polyacrylamide gel electrophoresis likely yield less inhibitory
> contaminants which would lead to cloned PCR products?  Will 3.0 or 4.4
> Kb fragments elute from the polyacrylamide during the purification
> process?

It has certainly been my (subjective) experience that DNA purified from
acrylamide is easier to clone than DNA from agarose, though I have never
performed a controlled experiment.

As for the purification itself - 3-4 kb is probably too big to elute by
passive diffusion, but I'm sure you could get it out by electroelution.

good luck,

          Julian Parkhill   -   J.Parkhill at bham.ac.uk
   CRC Laboratories, Inst. of Cancer Studies, Medical School,
 University of Birmingham, Edgbaston, Birmingham, B15 2TJ, U.K.
Disclaimer: If I thought anybody gave a damn about my opinions....

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