when I perform real-time PCR for different genes in human cancer cells, I usually got a base-line until about 25 cycles then linear amplifications starts. I use the following conditions:
1) cDNA concentration/well ~29 nanograms
2) forward or reverse primers concentration : 350 nM (final)
How can I shorten the length of the base-line and get amplifications earlier after about 15 cycles?
thanks in advance