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The optimized RAPD-PCR conditions, that can be utilized as a basic information for analysis of the genetic characteristics were developed for genetic analysis of four mulberry varieties, named Milsung, Chungil, Suil, and Hansung using a primer, OPY15 (5’-AGTCGCCCTT-3’) from Operon company. We tested several different factors for best PCR condition including concentrations of DNA, primer, MgCl₂, annealing temperature, number of PCR cycle, and presence/absence of pre-heating time at the begining of PCR reaction in the 25 μl volume. The best RAPD profiles were obtained using 50 ng of DNA, 1 μM of primer, 1.5 mM of MgCl₂, 45℃ of annealing temperature and an absence of pre-heating time. An establishment of the stable and reproducible RAPD-PCR conditions are expected to be useful for the subsequent RAPD-related investigation, such as genetic characterization of the mulberry varieties, re-establishment of phylogenetic relationships and development of new varieties.
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