how to improve the efficiency of my FQ-PCR - I can not get a beautiful amplification curve (Oct/12/2005 )

Hi,everyone!

I did the FQ-PCR in a long time, but my amplifcation curve still looks poor. I often find the good curve in the papers, but how can I do that also?

any suggestion thankful!

-big-boy-

I had tried primer 0.1~2.0pmol/ul.and Taqman probe 0.05~0.5pmol/ul. also changed the Ta vaule.
my PCR condition: 95℃ --1min; 95℃-10sec, 55℃-40sec, 40cycles.

thanks for any suggestion!

-big-boy-

QUOTE (big-boy @ Oct 12 2005, 01:23 AM)

I had tried primer 0.1~2.0pmol/ul.and Taqman probe 0.05~0.5pmol/ul. also changed the Ta vaule.
my PCR condition: 95℃ --1min; 95℃-10sec, 55℃-40sec, 40cycles.

thanks for any suggestion!

the best raw amplification curve is as follow:

-big-boy-

the best raw amplification curve is as follow:

-big-boy-