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Ethidium monoazide and propidium monoazide for elimination of unspecific DNA background in quantitative universal real-time PCR
Affiliation:1. Institute of Milk Hygiene, Milk Technology and Food Science, Department of Veterinary Public Health and Food Science, University of Veterinary Medicine, Veterinaerplatz 1, A-1210 Vienna, Austria;2. Clinic for Orthopaedics in Ungulates, Department of Small Animals and Horses, University of Veterinary Medicine, Veterinaerplatz 1, A-1210 Vienna, Austria;1. Department of Pharmacology, Institute of Biomedical Science, University of São Paulo (USP), 05508-000 São Paulo/SP, Brazil;2. Department of Physiology and Biophysics, Institute of Biomedical Science, University of São Paulo (USP), 05508-000 São Paulo/SP, Brazil;1. Psychology Department, Northeastern University, Boston, MA, United States;2. Genetics Department, Harvard Medical School, Boston, MA, United States;1. Department of Brain and Behavioral Sciences, University of Pavia, Via Forlanini 6, 27100 Pavia, Italy;2. Department of Biomedical Science, Sensory Neuroscience Group, Alfred Denny Building (B1 221), University of Sheffield, Western Bank, Sheffield S10 2TN, UK;1. School of Earth and Environmental Sciences, College of Natural Sciences, Seoul National University, Seoul 08826, Republic of Korea;2. Advanced Institutes of Convergence Technology, Suwon, Gyeonggi-do 16229, Republic of Korea;3. Department of Oceanography, College of Ocean Science and Technology, Kunsan National University, Kunsan 54150, Republic of Korea;4. Brain Korea Plus program, School of Earth and Environmental Sciences, College of Natural Sciences, Seoul National University, Seoul 08826, Republic of Korea;1. Department of Neurosurgery, Wuhan General Hospital of Guangzhou Military Command of Chinese PLA, Wuhan 430070, Hubei Province, China;2. Department of Neurosurgery, Tangdu Hospital, Fourth Military Medical University, No. 569 Xinsi Road, Baqiao District, Xi’an 710038, Shaanxi Province, China;3. Department of Aerospace Biodynamics, Fourth Military Medical University, Xi’an 710032, Shaanxi Province, China;4. Department of Neurology, Chinese PLA No. 451 Hospital, Xi’an 710054, Shaanxi Province, China;5. Department of Anesthesiology, Tangdu Hospital, Fourth Military Medical University, No. 569 Xinsi Road, Baqiao District, Xi’an 710038, Shaanxi Province, China
Abstract:Unspecific background DNA in quantitative universal real-time PCR utilizing a hydrolysis probe was completely suppressed by the addition of EMA or PMA to the PCR mix via cross-linking of the dyes to DNA during 650 W visible light exposure. The proposed procedure had no effect on the sensitivity of the real-time PCR reaction.
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