참외의 PCR 최적 조건 구명 = PCR Optimization in Cucumis melo

다국어 초록 (Multilingual Abstract)

This study was conducted to identify the optimized PCR condition in cucumis melo. Factorial experiments for template DNA concentration, MgCl2 concentration, amount of taq polymerase, and reaction temperature(denature, annealing, extension) were conducted. The best template DNA concentration was 40ng(0.5unit taq polymerase and 4.5mM MgCl2, and 1unit taq polymerase and 2.5mM MgCl2), 60ng(0.5unit taq polymerase and 2.5mM MgCl2, 0.5unit taq polymerase and 4.5mM MgCl2, 1unit taq polymerase and 2.5mM MgCl2, 1unit taq polymerase and 4.5mM MgCl2, and 1unit taq polymerase and 7.0mM MgCl2). The best MgCl2 concentration was 2.5mM(40ng template DNA and 1unit taq polymerase, 60ng template DNA and 0.5unit taq polymerase, and 60ng template DNA and 1unit taq polymerase)와 4.5mM(40ng template DNA and 0.5unit taq polymerase, 60ng template DNA and 0.5unit taq polymerase, and 60ng template DNA and 1unit taq polymerase), and 7.0mM(60ng template DNA and 1unit taq polymerase). The best amount of taq polymerase was 0.5unit(40ng template DNA and 4.5mM MgCl2, 60ng template DNA and 2.5mM MgCl2, and 60ng template DNA and 4.5mM MgCl2) and 1unit(40ng template DNA and 2.5mM MgCl2, 40ng template DNA and 4.5mM MgCl2, 40ng template DNA and 7.0mM MgCl2, 60ng template DNA and 2.5mM MgCl2, 60ng template DNA and 4.5mM MgCl2, and 60ng template DNA and 7.0mM MgCl2). When we consider results from template DNA concentration, MgCl2 concentration, and amount of taq polymerase, 6 conditions(60ng template DNA, 7.0mM MgCl2, 1unit taq polymerase; 60ng template DNA, 4.5mM MgCl2, 1unit taq polymerase; 60ng template DNA, 2.5mM MgCl2, 1unit taq polymerase; 60ng template DNA, 4.5mM MgCl2, 0.5unit taq polymerase; 60ng template DNA, 2.5mM MgCl2, 0.5unit taq polymerase; and 40ng template DNA, 2.5mM MgCl2, 1unit taq polymerase) were best combinations for the optimal PCR condition. Reaction temperatures for the optimal PCR condition were 90℃, 40℃, 72℃ and 92℃, 36℃, 72℃.

목차 (Table of Contents)

• 서언
• 재료 및 방법
• 1.식물재료
• 2.Genomic DNA 분리 및 quantification
• 3.PCR 조건
• 결과 및 고찰
• 1.Template DNA 농도
• 2.MgCl2 농도
• 3.Taq polymerase 농도
• 4.반응 온도