Comparison of three common DNA concentration measurement methods.
Accurate measurement of DNA concentration is important for DNA-based biological applications. DNA concentration is usually determined by the ultraviolet (UV) absorption, fluorescence staining, and diphenylamine reaction methods. However, the best method for quality assurance of measurements is unknown. Here, we comprehensively compared these methods using different types of samples. We found that all three methods accurately determined the concentrations of high-purity DNA solutions. After digestion of DNA samples, concentration measurements revealed that the PicoGreen dye method was very sensitive to the degradation of DNA. The three methods displayed different anti-jamming ability when contaminants such as transfer RNA (tRNA), protein, and organic chemicals were included in DNA solutions. The diphenylamine reaction method gave the highest accuracy, with an average error of approximately 10% between measured and true values. The PicoGreen dye method was influenced by tRNA and protein, and the UV absorption method was susceptible to all kinds of impurities. Overall, the diphenylamine reaction method gave the most accurate results when DNA was mixed with contaminants, the PicoGreen dye method was most suitable for degraded DNA samples or DNA extracted from processed products, and the UV absorbance method was best for evaluating the impurities in DNA solutions.
Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, People's Republic of China.
Xiaofei
X
Li
Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, People's Republic of China.
Yuhua
Y
Wu
Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, People's Republic of China.
Li
L
Zhang
Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, People's Republic of China.
Yinglong
Y
Cao
Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, People's Republic of China.
Yunjing
Y
Li
Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, People's Republic of China.
Jun
J
Li
Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, People's Republic of China.
Li
L
Zhu
Key Laboratory of Oil Crop Biology of the Ministry of Agriculture, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, People's Republic of China. Electronic address: wugang@caas.cn.
Gang
G
Wu
D018744:DNA, Plant / Q000032:analysis* / Q000737:chemistry; D004159:Diphenylamine / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D005470:Fluorometry; D009930:Organic Chemicals / Q000737:chemistry; D018515:Plant Leaves / Q000235:genetics; D010944:Plants / Q000235:genetics; D011506:Proteins / Q000737:chemistry; D012343:RNA, Transfer / Q000737:chemistry; D012639:Seeds / Q000235:genetics; D013056:Spectrophotometry, Ultraviolet
D018744:DNA, Plant; D005456:Fluorescent Dyes; D009930:Organic Chemicals; C099476:PicoGreen; D011506:Proteins; D012343:RNA, Transfer; D004159:Diphenylamine
Comparison; DNA concentration measurement; Diphenylamine reaction; PicoGreen dye; UV absorption