Absolute Quantification of References & Standards
Absolute quantification of references without a standard curve
Accurate genetic measurements often require comparison to reference samples and assay standards. Standardization of references is especially important in the field of metrology. For many organisms or applications, there is often no suitable reference sample available. Generation of reference standards using conventional real-time PCR requires consideration of how the reference sample will initially be calibrated, its long-term stability, and whether there is sufficient reference material for completion of all future studies. In addition, the lack of broadly adopted standards impacts comparison between laboratories.
Digital PCR does not rely on a reference sample or assay standard; it can be used for absolute quantification, measuring the exact copy number of a nucleic acid target of interest. This capability is especially useful for calibrating reference samples and assay standards when none exist. Through direct copy number determination, digitally measured assay standards can enable laboratories to compare results, with the assurance that measurements are based on the same absolute baseline.