Figure 5From: A Bayesian method for calculating real-time quantitative PCR calibration curves using absolute plasmid DNA standardsTo build the plasmid absolute standard construct, long oligonucleotides (~80 bp, Table 1) containing multiple primer binding sequences were designed such that their 3' ends overlapped. The two overlapping fragments were then combined into a single DNA molecule using overlap extension PCR [29]. Each partially overlapping fragment generated from the initial overlap extension was combined by a second overlap extension into a single full-length DNA construct.Back to article page