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Optimizing the generation of random amplified polymorphic DNAs in chrysanthemum
Authors:K Wolff  E D Schoen  J Peters-Van Rijn
Institution:(1) Department of Population Biology, University of Leiden, PO Box 9516, NL-2300 RA Leiden, The Netherlands;(2) Dept. of Biotechnology, TNO Nutrition and Food Research, PO Box 360, NL-3700 AJ Zeist, The Netherlands;(3) Centre for Applied Statistics, TNO Industrial Research, PO Box 155, NL-2600 AD Delft, The Netherlands
Abstract:Many conditions of the RAPD reaction procedure may influence the result. This paper presents rapid detection of influential factors with a fractional factorial experiment. A more extensive study of these factors is also presented. Polymerase brand, thermal cycler brand, annealing temperature, and primer, are important factors in obtaining good DNA yields and optimal fragment patterns. Each primer has its optimal annealing temperature, and this is not correlated with the GC content of the primer. Optimal species-primer combinations have to be found by trial and error.
Keywords:RAPDs  Experimental design  Fractional factorial  Chrysanthemum  PCR
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