PCR of AT-rich DNA - (Jun/09/2010 )
I have an extremely AT-rich sequence I have tried to amplify. I read the article by Su et al. (1996) "Reduced extension temperatures required for PCR amplification of extremely AT-rich DNA," and found that by reducing my extension temp to 60C, I do see more complete synthesis. However, I'm not sure how the reduction in temperature increases my product. Can someone please explain the mechanism behind this? Thanks.
-burroa03-
It has something to do with the stability of A-T bonds. They are less stable than G-C ones, and in higher temperatures the AT-rich sequence dissociates from the newly made double helix, thus making the creation of new strand difficult.
-Trof-