胡薇;陈宇;林来水;林思祖;

• 1:福建农林大学林学院
• 2:福建师范大学生命科学学院

摘要(Abstract)：

采用均匀设计,对影响黑木相思ISSR-PCR体系的引物、Mg2+、dNTP、Taq DNA聚合酶和模板DNA浓度等进行5因素5水平和5因素3水平两轮优化,建立了适合于黑木相思ISSR-PCR的体系:在20μL反应体系中,含引物0.30μmol.L-1,Mg2+3.00 mmol.L-1,dNTP 0.15 mmol.L-1,Taq DNA聚合酶0.75U,模板DNA 2.00 ng.μL-1.在此基础上对扩增程序中的循环次数、退火温度、退火时间进行筛选,获得的扩增程序为:94℃预变性5 min;接着进行33个循环:94℃变性35 s,52⁶1.5℃退火52 s,72℃延伸90 s;循环结束后,72℃延伸10 min.同时筛选得到10个扩增稳定、多态性丰富的ISSR引物.

关键词(KeyWords)： 黑木相思;均匀设计;ISSR-PCR

Abstract:

Keywords:

基金项目(Foundation): 福建省自然科学基金资助项目(B0410007)

作者(Authors): 胡薇;陈宇;林来水;林思祖;

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