OPTIMASI SUHU ANNEALING UNTUK EMPAT PRIMER RAPD PADA KACANG HIJAU (Vigna radiata L.)

H. Herman, Martupa Nainggolan, Dewi Indriyani Roslim
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引用次数: 1

Abstract

Determination of annealing temperature of the primer is the first step for genetic diversity analysis using molecular markers such as RAPD (Random Amplified Polymorphic DNA). This study aims to determine annealing temperature (Ta) of RAPD primers on Kampar Mungbean. Methods included total DNA extraction, electrophoresis, and annealing temperature optimization of four RAPD markers namely OPD-20, OPI-06, OPI-13, dan OPX-13. Optimization was conducted by reducing the Tm value (Time melting) of each primer with 3 (Tm-3) and 5 (Tm-5). The results showed that the optimization using OPD-20 and OPX-13 produced bands at Tm-3 and Tm-5. Meanwhile, optimization using OPI-06 and OPI-13 resulted in bands at Tm-3. The next step was to choose the exact Ta based on the clear and bright band. In conclusion, exact Ta for OPD-20, OPI-06, OPI-13, and OPX-13 were 36,1°C, 38,1°C, 35,4°C, and 32,5°C respectively.
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确定引物的退火温度是利用RAPD(随机扩增多态性DNA)等分子标记进行遗传多样性分析的第一步。本研究旨在确定甘帕绿豆RAPD引物的退火温度(Ta)。方法对OPD-20、OPI-06、OPI-13和OPX-13 4个RAPD标记进行总DNA提取、电泳和退火温度优化。以3 (Tm-3)和5 (Tm-5)降低每个引物的Tm值(Time melting)进行优化。结果表明,利用OPD-20和OPX-13进行优化后,在Tm-3和Tm-5处产生了条带。同时,使用OPI-06和OPI-13进行优化得到的波段位于Tm-3。下一步是根据清晰和明亮的波段选择准确的Ta。综上所述,OPD-20、OPI-06、OPI-13和OPX-13的精确Ta分别为36、1°C、38、1°C、35、4°C和32.5°C。
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