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.
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.