Usefulness of RAPD for genetic distance estimation in the European species Pinus sylvestris
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Random Amplified Polymorphic DNAs (RAPD) technique with single primers was tested for its usefulness in genetic distance estimation and population studies in the coniferous species Pinus sylvestris. DNA markers allow for direct analysis both coding and noncoding regions of the genome. The technique of detection DNA variations using RAPD markers has become a popular tool in genetic studies. Different reaction conditions were tested in order to get the optimal resolution of fragments, specificity and reproducibility of patterns. In this preliminary investigation, a high MgCl2 concentration (5,5 mM) together with a low primer concentration (0,2 M) in the polymerase chain reaction (PCR) mixture yielded the best amplification products. Amplified fragments were scored as the presence or absence fragments.
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I. Szyp-Borowska, Zaklad Fizjologii i Genetyki Drzew Lesnych, Instytut Badawczy Lesnictwa, ul. Bitwy Warszawskiej 1920 nr 3, 00-973 Warszawa, Poland