为确保光核桃ISSR反应条件的稳定性,以改良的CTAB法提取光核桃基因组DNA,对模板DNA浓度、MgCl2浓度、dNTPs浓度、Taq酶用量、引物浓度等因素进行了优化,确立了光核桃ISSR反应体系。结果表明:最佳反应体系为模板浓度40ng/μL、引物浓度0.4μmol/L、Mg2+的浓度2.5mmol/L、酶浓度0.5U、dNTPs浓度0.5mmol/L,为光核桃这一优质的种质资源的遗传多样性研究奠定了理论基础。
To ensure the stability of ISSR for Prunus mira koehne Kov et.Kpst,total genomic DNA was extracted by improved CTAB method,the factors including template DNA concentration,MgCl2 concentration,dNTPs concentration,Taq polymorphism dose and primer concentration were studied to establish optimized ISSR reaction system.The result showed that template concentration 40ng/μL,primer concentration 0.4μmol/L,Mg2+concentration 2.5 mmol/L,enzyme concentration 0.5U,dNTPs concentration 0.5mmol/L were the optimized reaction system.The study would provide the foundation for the genetic diversity on germplasm resources of Prunus mira koehne Kov et.Kpst.