Optimization of ISSR-PCR Amplification in Xanthoceras sorbifolia Bunge

doi:10.11924/j.issn.1000-6850.2013-2119

Abstract

Abstract: The aims were to establish an optimal reaction system of ISSR-PCR in Xanthoceras sorbifolia Bunge, and provide theoretical basis for its genetic diversity research. From X. sorbifolia artificial forest in Chankou forest farm of Dingxi City, picking health and tender leaves as experiment material, the concentrations of main factors influencing the effects of ISSR-PCR were tested by single factor experiments respectively, the tested factors including template DNA concentration, annealing temperature, dNTP concentration, template DNA concentration, annealing temperature, Mg²⁺ concentration, primer concentration and Taq DNA polymerase. Through test, this study had found suitable conditions for each factor, established and optimized X. sorbifolia’s reaction system of ISSR-PCR, namely the optimal reaction system for ISSR analysis in X.sorbifolia, included 30 ng template DNA, 2.5 mmol/L Mg2 + , 0.2μmol/L primer, 0.20 mmol/L dNTP and 1 U of Taq DNA polymerase in a total volume of 20μL. The study indicated that atlas with fine diversity and clear strap was gained.

Lu Juan, Chai Chunshan, Wu Wenjun, Qi Jianli. Optimization of ISSR-PCR Amplification in Xanthoceras sorbifolia Bunge[J]. Chinese Agricultural Science Bulletin, 2014, 30(1): 32-36.

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