[ Objective] The aim was to establish the optimal ISSR-PCR system for Armeniaca sibirica. [ Method] With Armeniaca sibirica as experimental materi- al, genomic DNA was extracted from tender leaves using CTAB method. O...[ Objective] The aim was to establish the optimal ISSR-PCR system for Armeniaca sibirica. [ Method] With Armeniaca sibirica as experimental materi- al, genomic DNA was extracted from tender leaves using CTAB method. Orthogonal experiment L9 （34） was conducted with four factors （Mg2＋ , dNTPs, primers, Taq DNA polymerase） in ISSR-PCR system ofArmeniaca sibirica at three levels. The results of orthogonal experimental design were analyzed by using SPSS statisti- cal software. [Result] Different levels of various factors all had significant effects on PCR results, specifically, the amount of Taq DNA polymerase had the most significant effect. The optimal ISSR-PCR reaction system of Armeniaca sibirica was established ： the total PCR reaction volume was 20 ~1, containing 1.0 U of Taq DNA polymerase, 1.0 mmol/L MgCI2,0.15 mmol/L dNTPs and 0.30 ~mol/L primers. Validation result shows that this reaction system is very stable and strongly operational. [ Conclusion] This study laid the foundation for genetic diversity analysis and germplasm resource investigation of Arnniaca sibirica.展开更多
基金Supported by Start-up Fund for High-level Scientific Research Personnel(11GK30)
文摘[ Objective] The aim was to establish the optimal ISSR-PCR system for Armeniaca sibirica. [ Method] With Armeniaca sibirica as experimental materi- al, genomic DNA was extracted from tender leaves using CTAB method. Orthogonal experiment L9 （34） was conducted with four factors （Mg2＋ , dNTPs, primers, Taq DNA polymerase） in ISSR-PCR system ofArmeniaca sibirica at three levels. The results of orthogonal experimental design were analyzed by using SPSS statisti- cal software. [Result] Different levels of various factors all had significant effects on PCR results, specifically, the amount of Taq DNA polymerase had the most significant effect. The optimal ISSR-PCR reaction system of Armeniaca sibirica was established ： the total PCR reaction volume was 20 ~1, containing 1.0 U of Taq DNA polymerase, 1.0 mmol/L MgCI2,0.15 mmol/L dNTPs and 0.30 ~mol/L primers. Validation result shows that this reaction system is very stable and strongly operational. [ Conclusion] This study laid the foundation for genetic diversity analysis and germplasm resource investigation of Arnniaca sibirica.
