Comparison of Methods for Accurate Quantification of DNA Mass Concentration with Traceability to the International System of Units

Citations Over TimeTop 10% of 2010 papers

Abstract

Accurate estimation of total DNA concentration (mass concentration, e.g., ng/muL) that is traceable to the International System of Units (SI) is a crucial starting point for improving reproducible measurements in many applications involving nucleic acid testing and requires a DNA reference material which has been certified for its total DNA concentration. In this study, the concentrations of six different lambda DNA preparations were determined using different measurement platforms: UV Absorbance at 260 nm (A(260)) with and without prior sodium hydroxide (NaOH) treatment of the DNA, PicoGreen assay, and digital polymerase chain reaction (dPCR). DNA concentration estimates by A(260) with and without prior NaOH treatment were significantly different for five of the six samples tested. There were no significant differences in concentration estimates based on A(260) with prior NaOH treatment, PicoGreen analysis, and dPCR for two of the three samples tested using dPCR. Since the measurand in dPCR is amount (copy number) concentration (copies/muL), the results suggest that accurate estimation of DNA mass concentration based on copy number concentration is achievable provided the DNA is fully characterized and in the double-stranded form or amplification is designed to be initiated from only one of the two complementary strands.

Related Papers

• → Nondestructive Detection of Water Stress in Tomato Plants by NIR Spectroscopy.(2001)5 cited
• → The Linear Relationship between Concentrations and UV Absorbance of Nitrobenzene(2014)1 cited
• → Direct Spectrophotometric Determination of Iron in the HCl Soluble Fraction of Carbonate Rocks(1966)
• → Effect of Inorganic Salts on the Determination of Serum Alubmin by Dye-Binding Method with Absorbance Decrease(2007)
• Infrared Absorbance of SiN_x and SiC(2006)