i use pcr product as template in real time pcr to get the abslute stundard curve and then calculate the amount of my samples(genomic dna). always i have band for my samples but i have no band for stundards! but specific tm and peak for both!!
the starting point of diluation serial have E7 copy number of target.
another problem is high efficiency in stundard curev or low effiinecy ! i can't get a good result !
anyone know if there is any diffrence in pcr condition between genomic dna and pcr product!
thaks for any help