毛白杨ISSR反应体系的建立及优化

基金项目:

国家自然科学基金项目(30571516)

详细信息

Establishment and optimization of an ISSR reaction system for Populus tomentosa Carr

1. Key Laboratory for Genetic and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, Beijing Forestry University, 100083, P. R. China;
2. Department of Biotechnology, Beijing Agricultural College, 102206, P. R. China;
3. Guanxian County Tree Nursery, Shandong Province, 252528, P. R. China

摘要: 为应用ISSR技术开展毛白杨遗传变异分析、辅助育种、品种鉴定、系统进化等研究,以(GTG)₆为引物,通过单因子实验分别研究了退火温度、引物浓度d、NTP浓度、Mg²⁺浓度、Taq DNA聚合酶用量对毛白杨ISSR-PCR反应的影响,建立并优化了适宜于毛白杨ISSR分析的扩增体系:20μL PCR反应体系,1×Taq DNA酶缓冲液(10 mmol/L Tris-HCl,50 mmol/L KCl,0.1%Trion X-100,pH 9.0),1.5 mmol/L MgCl₂,1.0 U Taq酶,10 ng模板DNA,0.2μmol/L引物,0.2 mmol/L dNTP.引物(GTG)₆的最适退火温度为61℃.
- 毛白杨 /
- ISSR /
- PCR /
- 条件优化
Abstract: In order to use ISSR markers to study genetic variation,assistant breeding,cultivar identification and phylogenesis of Populus tomentosa,the annealing temperature,concentration of primer,dNTP,Mg²⁺ and Taq DNA polymerase dosage on ISSR-PCR amplications were tested to determine their optimal levels and establish the following optimal reaction system for ISSR analysis in P.tomentosa.The results showed that the optimal conditions for ISSR-PCR of P.tomentosa were as follows:PCR reaction volume of 20 μL,1×Taq buffer(10 mmol/L Tris-HCl,50 mmol/L KCl,0.1% Triton X-100,pH 9.0),1.0 U Taq DNA Polymerase,0.2 μmol/L primer,0.2 mmol/L dNTP,1.5 mmol/L MgCl-2 and 10 ng template DNA.The optimized annealing temperature was 61℃ for primer(GTG)-6.
- Populus tomentosa /
- ISSR /
- PCR /
- optimized conditions

田中君，肖人源，李巍，沈阳，童水光. 干燥介质旁通及回热对闭式热泵干燥系统除湿率影响的分析. 工业加热. 2019(02): 25-29 .

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