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Position: Home > Articles > Optimization of ISSR Reaction System for Species of Loquat (Eriobotrya japonica Lindl.cv) Chinese Agricultural Science Bulletin 2010,26 (7) 26-30

枇杷ISSR扩增反应体系的优化

作  者:
赵依杰;王江波;张小红;施维属;林航
单  位:
福州市农业科学研究所;福建农林大学园艺学院
关键词:
枇杷;ISSR;正交设计;体系优化
摘  要:
采用正交设计对影响枇杷ISSR-PCR的模板DNA、dNTPs、Primer、Taq酶及Buffer(含Mg2+)进行优化。试验结果表明:20μL反应体系中,含模板DNA5ng、dNTPs0.25mmol/L、Primer0.25μmol/L、Taq酶0.5U和10×Buffer(含Mg2+)3μL。此外,从95条ISSR引物中筛选出11条具有丰富的多态性位点的引物,同时优化各引物退火温度。
译  名:
Optimization of ISSR Reaction System for Species of Loquat (Eriobotrya japonica Lindl.cv)
作  者:
Zhao Yijie1,Wang Jiangbo2, Zhang Xiaohong1, Shi Weishu2, Lin Hang1 (1Fuzhou Institute of Agricultural Sciences, Fuzhou 350018; 2College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002)
关键词:
Loquat (E.japonica Lindl.cv ); ISSR; orthogonal design; system optimization
摘  要:
The factors of template DNA, dNTPs, Primer, Taq polymerase and Buffer (containing Mg2+), greatly influencing ISSR-PCR of loquat, were optimized by orthogonal design. The result shows that the optimum amplification condition of ISSR-PCR is DNA template 5 ng, dNTPs 0.25 mmol/L, Primer 0.25 μmol/L,Taq Polymerase 0.5 U and 10×Buffer (contain Mg2+) 3 μ in 20 μL reaction volumes. Besides, 11 ISSR primers has been obtained with high stability and polymorphism simultaneity obtained for ISSR-PCR annealing temperature from 95 ISSR primers.

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