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Selection of RAPD Reaction Conditions for Genus Dendranthema

Dai Silan; Chen Junyu; Li Wenbin(College of Landscape Architecture, BFU 100083)(Institute of Genetics, Academia Sinica, Beijing 100101)  
Many factors of the RAPD reaction procedure may influence the results. This paper presents selected RAPD reaction condition for analysis of genetic relationship among Dendranthema species. A more extensive study of influential factors is also presented. To get good results, the reaction should be performed in a volume of 20μl containing 10mmol/lTris-HCl(pH8. 3), KCI 50mmol/l, MgCl2 2mmol/l, gelatin 0. 001%, dNTP 200μmol/l,primer 50 ̄200ng, Taq DNA polymerase 0. 75 ̄1. 0 unit, genomic DNA 2 ̄10ng. The mixture was laid on thermal cycle. After 3 minutes at 92C, 40 cycles were run. Each cycle consisted of 45 seconds at 92℃, 1 minute at 50℃, 2 minutes at 72℃. Then 10 minutes at 72℃.Plant growth state, methods of DNA isolation were also discussed as influential factors.
【Fund】: 国家自然科学基金
【CateGory Index】: S682.110.2
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