摘要
[目的]确定臭椿SRAP-PCR反应条件,为进一步研究臭椿SRAP分子标记提供依据。[方法]以新疆吐鲁番3号和江西臭椿叶片DNA为材料,利用引物组合EM1-EM8进行SRAP-PCR反应的L16(45)正交试验,建立了臭椿SRAP-PCR反应体系,新复极差法对体系进行方差分析,并对体系的稳定性进行检测。[结果]确定臭椿SRAP-PCR反应体系为:模板DNA 2.5 ng/μl、Mg2+1.75 mmol/μl、dNTPs0.3 mmol/μl、Taq酶0.3 U/μl、引物0.6μmol/μl、10×PCR Buffer 2.5μl;该体系稳定,适用于臭椿的SRAP反应。[结论]该试验优化的SRAP反应体系,将为臭椿种质资源多样性评价、分子标记,以及遗传连锁图谱构建奠定基础。
[Objective] The aim was to confirm the SRAP-PCR reaction conditions of A.altissima,to provide basis for further studying SRAP markers of A.altissima.[Methods] L16（45）orthogonal test of SRAP-PCR reaction was carried out using primers combination EM1-ME8 based on the materials of Xinjiang Turpan No.3 and blades DNA of Jiangxi A.altissima,moreover,established SRAP-PCR reaction system of A.altissima and new multiple range analysis of variance on the system,and the stability of the system was detected.[Result] The SRAP-PCR reaction system of A.altissima is determined as follows：templates DNA 2.5 ng/μl,Mg2＋ 1.75 mmol/μl,dNTPs 0.3 mmol/μl,Taq enzyme 0.3 U/μl,primers 0.6 ummol/μl,10×PCR Buffer 2.5 μl.The system is stable and adaptable for SRAP reaction of A.altissima.[Conclusion] The optimized SRAP reaction system will provide basis for the evaluation of genetic diversities,molecule marker assisted breeding and linkage mapping of A.altissima.
出处
《安徽农业科学》
CAS
2012年第5期2570-2573,共4页
Journal of Anhui Agricultural Sciences
基金
江苏省林业三新工程LYSX(2009)39
关键词
臭椿
SRAP-PCR
体系优化
正交设计
Ailanthus altissima
SRAP-PCR
System optimization
Orthogonal design
