We tried to measure the telomere length of T cells by using qPCR method. The sequence of the primers was listed in the bottom. Serial dilutions of genomic DNA was created to generate a standard Ct value curve for the assay. But I found the amplification efficiency of Telo was about 109%, 100% for single copy genes (SCG). My point is that the amplification efficiency of Telo doesn't need to be 100% due to the varying length of telomere length, and the T/S ration is the ratio of CT value of Telo to the CT value of SCG. I don't know if i am right.

Thank u !

Telo 1: 5'-CGGTTTGTTTGGGTTTGGGTTTGGGTT-TGGGTTTGGGTT-3'

Telo 2: 5'-GGCTTGCCTTACCCTTACCCTTACCCTTACCCTT-ACCCT-3'

SCG 1: 5'-GCTTCTGACACAACTGTGTTCACTAGC-3'

SCG 2: 5'-CACCAACTTCATCCACGTTC-ACC-3'