Affiliation:
1. National Institute of Metrology
2. Beijing University of Chemical Technology
Abstract
For the sake of accurate quantification, it was investigated the factors affecting the Picogreen measurement on genomic DNA of legionella pneumophila. As a binding and excitation working manner, PicoGreen fluorescent signal intensity showed to be dependent on incubation time, environmental pH and temperature, nuclear acid fragment length and concentration. Based on the optimized detection conditions, we carried out the methodology investigation including linear dependence, repeatability, LOD and LOQ. From the data, it can be concluded that PicoGreen can be used as a rapid and sensitive candadite characterization method to quantify total DNA of legionella pneumophila.
Publisher
Trans Tech Publications, Ltd.
Reference7 articles.
1. Lu Shengdong, Experimental Techno logy of Modern Molecular Biology, 2nd edition, Peking Union Medical College Press 1999. 69.
2. Ausubel F. M., Quantitation of DNA and RNA with absorption and fluorescence spectroscopy. Current Protocols in Molecular Biology; Ed., John Wiley & Sons, Inc.: New York, 2001; Appendix 3D.
3. Singer, V. L., Jones, L. J., Yue, S. T., Haugland, R. P. , Characterization of PicoGreen reagent and development of a fluorescence-based solution assay for double stranded DNA quantitation. Anal. Biochem. 1997, 249, 223–238.
4. Barman SP, Lunsford L, Chambers P, et a l. Two method s for quantifying DNA extracted from poly ( lactide - co - glycolide) microspheres. J Con trolled Re lease, 2000, 69 ( 3) : 337.
5. Pharmacopoeia Commission, Pharmacopoeia of the People's Republic of China,2010 edition,China Medical Science and Technology Press,(2010).
Cited by
1 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献