本研究以玉米自交系(Mo17)為材料,以OPERONA-18為核酸引子(primer),利用快速省時之空氣溫度循環裝置(Air Thermo-Cycler)進行聚合酵素連鎖反應(Polymerase Chain Reaction, PCR),希望建立玉米多條帶(band)圖譜之核酸逢機增殖多型性(Random Amplified Polymorphic DNA, RAPD)分析系統,以便進行後續之玉米種原遺傳歧異分析。試驗結果認為Mg(上標 ++)濃度為3mM、核酸引子濃度為4μM、融合(annealing)溫度為36℃、融合時間為0秒、變溫循環45次為進行分析之最適條件,且採此分析條件成功地以核酸引子OPERON A-02於7個玉米自交系(B57、B76、H95、HCBSA、Mo17、R98-13、朴1343)間建立了清晰的多條帶圖譜而偵得彼此間之差異。
The objective of this study is to optimize Random Amplified Polymorphic DNA (RAPD) fingerprinting analysis system of maize. Various conditions of polymerase chain reaction were tested in a thermo-cycling instrument using maize inbred line Mo17 and a tenbase random primer. The amplification condition optimized for the number of bands was 3mM Mg(superscript ++), 4μM primer, annealing extending and denaturing at 36℃’ 72℃ 94℃ for 0、45 and 0 second, respectively, for 45 cycles. Differences were detected among maize inbred lines B57、B76、H95、HCBSA、Mo17、R98-13 and Ds1343 with primer OPERON A-02.