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A Study of Optium Condition of RAPD for the Analysis of Genetic Characteristics by Autumn Leaf Color of Zelkova serrata  

Choi, Byoung Kon (Department of Environmental Resources, Sang Myung Univ.)
Bang, Kwang Ja (Department of Environmental Resources, Sang Myung Univ.)
Publication Information
Journal of the Korean Society of Environmental Restoration Technology / v.7, no.5, 2004 , pp. 94-99 More about this Journal
Abstract
This study was carried out to find out what is the optimum conditions for RAPD of Zelkova serata. We changes the factors what affect to PCR band patterns, as a result, we established the optimum conditions as follows; template DNA 100mg, Primer 0.25uM, dNTP 100mM, Taq polymerase 1.0u, and total reaction volume was filled up to 10uL with distilled water. As the amount of primers went higher, PCR reaction rates were lowered. This reason was cause by exhaustion of primers during initial reaction. The amount of dNTP didn't showed noticable differtations between the range, but the optimum amount was 100mM for efficiency. Taq polymerase 1.0 unit was the best in the range. As the concentration of polymerase were increased, many non-specific bands were appeared, In primer selection, most Openron Random Primers are amplified in this experiment. The primers GC contents were 60, and set A, B, C, D, E, X were tested. Thermal cycler(ASTEC PC808, Japan) condition was, $95^{\circ}C$, 5min, initial denaturation, $94^{\circ}C$, 20sec, denaturation, $37^{\circ}C$, 40sec, annealing, $72^{\circ}C$, 1min, extention, 45cycle repeated and final extention $72^{\circ}C$10min.
Keywords
Zelkova; PCR condition; primer selection;
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