I want to determine the amount of expression by performing qPCR of a bacterial gene region. For this, I compared some primers found in the literature. Some of these primers targeted the starting part of the gene, while some expressed the last part of the gene. Does it matter where the primer is linked to the gene to determine the expression level by making qpcr? When making qpcr, does the binding of the primer from the end or beginning to the gene site affect the expression level?