毛白杨ISSR反应体系的建立及优化

被引：25

作者：

冯夏莲 ^{[1
]}

何承忠 ^{[1
]}

张志毅 ^{[1
]}

安新民 ^{[1
]}

杨凯 ^{[2
]}

张有慧 ^{[3
]}

机构：

[1] 北京林业大学林木花卉遗传育种教育部重点实验室

[2] 北京农学院生物技术系

[3] 山东省冠县苗圃

来源：

北京林业大学学报 | 2006年 / 03期

关键词：

毛白杨; ISSR; PCR; 条件优化;

D O I：

10.13332/j.1000-1522.2006.03.011

中图分类号：

S792.117 [毛白杨];

学科分类号：

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℃.

引用

页码：61 / 65

页数：5

共 28 条

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