question about pcr - (Sep/04/2010 )

dear all

I working on goat and sheep promotor gene amplification.
I have a high quality DNA and I extracted it from blood.
One day I'll get the answer, but the next day with the same protocol does not get the answer in pcr
In the case I do many expriment but dont different in results.
please help me emergency

Hi iraj,

You should check the following items.

1)Verify that you have not forgetten any reagents in your master mix.
2)PCR protocol?
3)Change dNTPs.
4)Check your primers for dimerization errors or hairpin formation.
5)DNA quality?

Did you use the same sample?

cagdas on Sun Sep 5 18:11:31 2010 said:

Hi iraj,

Did you change all the reagents?
How many samples do you analyze in one go?
What' s your pcr reaction condition, protocol and expected amplicon size?
Do you run your fragments out on acrylamide or agarose?
Is agarose at the correct % for the fragment sizes you are looking at?