Polymerase - (Oct/31/2007 )

Hello,
Which is the best polymerase to amplify STR sequence in rice?, because I have used gotaq and the results are not stable, sometimes there is amplification and sometimes no amplification, we have used the same STR.

Regards

Hi there,

If you have amplification sometimes and sometimes not, I very much doubt that it is the polymerase that is the problem. If you give us more info about your PCR, someone may be able to give you some suggestions on what could be going wrong.

Laura

Hello
thank you, I have changed gotaq by prime star, so it works well with STR.
Regards

from the sounds of this, it might be that your template is not clean enough. When there is a PCR problem, the first suspect is the DNA template as it is usually at fault.

Is it possible to clean up the DNA any further? If not, consider making dilutions of your template. Try dilutions of 1:20 to 1:50. This dilutes away the contaminants in the DNA, and PCR amplifies the signal, so you still see something on your gels.