PCR getting smear

[Paul Brian Scruggs]

Zahra Fayazi <zahra.fayazi at jcu.edu.au> wrote:

>Hello everyone
>In order to amplyfy a 420 bp fragment from mice spleen cells infected 
>with Brucella suis, i applied different procedures, but with all of them 
>I have only got smear instead of band. The procedures which I applied 
>rang from phenol extraction to just lysis of cells and some teatment in 
>order to get inhibitors for PCR. Please if any one can give me advise on 
>how to get ride of this smear and have sharp band, I will be very 
>grateful to hear from.

Zahra,

If all else fails, you can try nested (internal) primers and run a
secondary PCR.

Brian Scruggs