Limitations of methods for measuring the concentration of human genomic DNA and oligonucleotide samples

We compared different methods (absorbance, fluorescent dye-binding, and digital PCR) for measuring the concentrations of human genomicDNA from cultured cells and absorbance measurements of a synthetic DNAoligonucleotide. NISTStandard Reference Material (SRM) 2082, a pathlength absorbance standard, was used to benchmark the absorbance measurements done with microvolume spectrophotometers and a microvolume plate reader. Control absorbance values were measured on a high accuracy spectrophotometer and a NISTcalibrated pathlength cuvette. Measurements of the human genomicDNA sample were done with several types of fluorescent dye binding assays using different DNA calibrators. The fluorescent dye binding methods gave different results for genomic DNAdepending upon the type of DNA calibrator and the fluorescent dye that was used. The human genomicDNA sample was also characterized by using six different droplet digital PCR assays ( ampliconslocated on different chromosomes) to measure the average copy numb...