白桦ISSR-PCR反应体系的优化

被引：26

作者：

杨传平

潘华

魏志刚

姜静

机构：

[1] 东北林业大学

来源：

东北林业大学学报 | 2005年 / 06期

关键词：

白桦; ISSR标记; 反应体系;

D O I：

10.13759/j.cnki.dlxb.2005.06.001

中图分类号：

S792.153 [白桦];

学科分类号：

0829 ; 0907 ;

摘要：

以白桦基因组DNA为模板,研究了ISSR的影响因素,建立一套适宜于白桦的ISSR优化反应体系及程序。即20μL反应体系中,Mg2+浓度范围为0.5～3.0mmol·L-1,dNTPs浓度范围为0.10～0.30mmol·L-1,Taq聚合酶浓度范围为1.0～2.0U,模板DNA浓度为30～120ng,同时含有RNA的模板DNA对ISSR扩增也略有影响,引物浓度范围为0.2～0.4μmmol·L-1。通过梯度PCR测验,确定了适宜的退火温度为49℃。反应程序为:第一个循环基因组DNA经94℃预变性5min;30个循环为94℃变性30s,49℃退火30s,72℃延伸30s;最后一个循环完成后,在72℃延伸7min。

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页码：1 / 3

页数：3

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