Q-PCR and noisy melting curves

[cc4280]

Hi,

I'm currently using Q-PCR to look at changes in gene expression under different experimental conditions. However, for one of the primers I'm using, I get a very noisy baseline. As expected, the noise increases as the primer concentration is reduced, but I don't understand why the noise is so high when the Ct value is still quite low. Are there any factors that contribute to a nosiy baseline? contaminations in the Q-PCR mixture perhaps?

 

Any help would be greatly appreciated.

[troy]

This is an old post but seems like a good place for my first post.

 

I had this happen to me. I did a number of things to resolve the problem. First I clean my primers. Then I personally wash all tips/tubes/plates etc (by plates I don't mean the 384 well one only 96). And I used Q-PCR mixtures that only I had used (can't trust everyone else is as careful as you may be). I used Milli-Q water and if I didn't get new water and storage bottle regularly there tended to be more noise due to contamination.

 

Hope this is some help.