DNA degradation

Dr. Duncan Clark Duncan at nospam.demon.co.uk
Fri Oct 15 03:12:20 EST 1999

In article <VNsN3.2182$u32.200719 at WReNphoon4>, Claudia Vorbach
<cvorbach at howard.genetics.utah.edu> writes
>my preped targeting vector (22kb) gets degradet during restriction enzyme
>digest, or even just with different enzyme buffers alone. Did several preps,
>some were ok until the problem started. exchanged all solutions, same
>degradation occurs. Other plasmids preped during the last weeks behave just
>fine. Tried to clean DNA prep with Phenol or different columns, nothing
>helps !!! Does anybody have an idea why my DNA gets degraded and how I can
>prevent it ???? THANX

The above is a bit vague to fault find on.

What is the source of the vector, as in strain?

What is your normal purification procedure?

What is the nature of the degradation, smear, discrete bands etc?

What RE's are you using?

The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
DNAmp Ltd.
Tel: +44(0)1252376288
FAX: +44(0)8701640382

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