A simple and fast method for cloning and analyzing polymerase chain reaction products |
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| Affiliation: | 1. From the Fishberg Center for Neurobiology, Mount Sinai School of Medicine, New York City, New York, USA;2. Psychiatry Department, Mount Sinai School of Medicine, New York City, New York, USA;1. Department of Plant Biology and Soil Science, University of Vigo, Campus Lagoas-Marcosende, 36310- Vigo, Spain;2. International Center for Biosaline Agriculture (ICBA), P.O. Box 14660, Dubai, UAE;1. Department of Neurology, the Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan, China;2. Department of Ultrasound, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan, China;3. Department of Neurology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan, China;1. Departamento de Ingeniería Química y Biotecnología, Facultad de Ciencias Físicas y Matemáticas, Universidad de Chile, Beauchef 850, Santiago, Chile;2. Instituto de Quimica, Universidade Federal do Rio Grande do Sul, Avenida Bento Goncalves 9500, Porto Alegre, Brazil;1. Department of Robotics and Mechatronics, AGH University of Science and Technology, Al. Mickiewicza 30, 30-059 Krakow, Poland;2. Institute of Mathematics, Jagiellonian University, ul. Prof. Stanisława Łojasiewicza 6, 30-348 Krakow, Poland;1. Molecular Imaging Branch, National Institute of Mental Health, Bethesda, MD, USA;2. Office of the Clinical Director, National Institute of Mental Health, Bethesda, MD, USA;3. Memory Disorders Program, Georgetown University, Washington, DC, USA |
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| Abstract: | A method describing a fast and efficient way for cloning polymerase chain reaction (PCR) products is presented that involves end repair and purification of the PCR product, followed by kinasing and ligation to the vector with the use of a temperature gradient. Efficiency of ligation was estimated to be 50%–70%. Following transformation, cells are plated on MacConkey agar. Bacteria from selected colonies are used directly from the plates for screening without any subsequent purification. Using this protocol, PCR products can be efficiently cloned quickly and economically. |
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