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Optimization for ISSR Reaction System of Hordeum vulgare subsp.Vulgare with Orthogonal Design

Tao Zhang

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Abstract

Five factors(Taq polymerase,Mg2+,dNTP,primer,DNA template) of Hordeum vulgare subsp.Vulgare(barley) ISSR amplification system were optimized using ISSR reaction system to analyze the evolutional relationship of barley.And then the optimum reaction system which contained 0.5 U Taq polymerase,1 μmol/L Mg2+,0.2 μmol/L dNTP,0.3 μmol/L primer,50 ng DNA template in a total volume of 25 μl amplification system was established.The optimum reaction system contributed greatly to use the ISSR-PCR technology to classify the quality of Hordeum vulgare subsp.Vulgare and mine the new genes.

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What this paper is about

Five factors(Taq polymerase,Mg2+,dNTP,primer,DNA template) of Hordeum vulgare subsp.Vulgare(barley) ISSR amplification system were optimized using ISSR reaction system to analyze the evolutional relationship of barley.And then the optimum reaction system which contained 0.5 U Taq polymerase,1 μmol/L Mg2+,0.2 μmol/L dNTP,0.3 μmol/L primer,50 ng DNA template in a total volume of 25 μl amplification system was established.The optimum reaction system contributed greatly to use the ISSR-PCR technology to classify the quality of Hordeum vulgare subsp.Vulgare and mine the new genes.

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Available abstract

Five factors(Taq polymerase,Mg2+,dNTP,primer,DNA template) of Hordeum vulgare subsp.Vulgare(barley) ISSR amplification system were optimized using ISSR reaction system to analyze the evolutional relationship of barley.And then the optimum reaction system which contained 0.5 U Taq polymerase,1 μmol/L Mg2+,0.2 μmol/L dNTP,0.3 μmol/L primer,50 ng DNA template in a total volume of 25 μl amplification system was established.The optimum reaction system contributed greatly to use the ISSR-PCR technology to classify the quality of Hordeum vulgare subsp.Vulgare and mine the new genes.

Key concepts: Hordeum vulgare, Primer (cosmetics), System optimization, Taq polymerase, Polymerase chain reaction, Biology, Botany, Chemistry

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