Comparison of six DNA quantification methods

Abstract

Six commercial preparations of human genomic DNA were quantified using six quantification methods including UV spectrometry, SYBR-green dye staining, slotblot hybridization with the probe D17Z1, and three TaqMan real-time PCR assays: Quantifiler™ Human DNA Quantification kit, Quantifiler™ Y DNA Quantification kit, and RB1 rt-PCR. In general, all methods measured higher DNA concentrations than expected based on the information by the suppliers of the human DNA preparations. The Quantifiler™ Human DNA Quantification kit gave the highest measures of the DNA concentrations of five of the six human DNA preparations compared to the other five quantification methods. When the Quantifiler™ human DNA standard was replaced by a different commercial human DNA preparation (G147A, Promega) to generate the DNA standard curve in the Quantifiler™ Human DNA Quantification kit, the DNA quantification results of the human DNA preparations were comparable to those of the other DNA quantification methods. The results indicate a calibration problem with the Quantifiler™ human DNA standard for its use with the Quantifiler™ Human DNA Quantification kit. The possible reasons of the problem are discussed, and a solution is suggested. The results emphasise the need for standard reference DNA material and standard methods for DNA quantification.