梨ISSR-PCR反应体系的正交优化研究
On Optimization of Reaction System in Pear-Using Orthogonal Design
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摘要: Orthogonal design has been applied to optimize ISSR‐PCR amplification system of in pear in five factors such as primer ,dNTPs ,template DNA ,Taq DNA polymerase and Mg2+ at four levels .The result of PCR has been analyzed by range analysis and variance analysis .An optimal reaction system (25μL) has been established ,i .e .2 .5 μL 10 × Buffer ,2 .0 mmol/L M g2+ ,0 .25 mmol/L dN T Ps ,0 .25 μmol/L prim‐er ,60 ng template DNA ,0 .75 U Taq DNA polymerase .The results indicate that the optimized reaction system was stable and repeatable .The objective of the study is to provide a scientific basis for the con‐structing DNA fingerprints and genetic diversity analysis of in pear .Abstract: 梨; 正交设计; ISSR体系优化
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