山楂ISSR分析体系的建立和优化Establishment and optimization of ISSR system in Crataegus spp.
代红艳;张志宏;周传生;李贺;郭修武;
摘要(Abstract)：
为在DNA分子水平上对山楂种质资源进行分析奠定基础,对退火温度、循环次数、DNA模板质量、Mg2+浓度、Taq DNA聚合酶的种类和浓度等影响ISSR反应的因素进行研究,建立并优化了山楂的ISSR分析体系。优化的山楂ISSR分析体系为:采用改进的CTAB法或QIAGEN试剂盒法提取总DNA;PCR反应体积为20μL,内含1×PCR反应缓冲液(Promega公司),3.0mmol/L MgCl2,0.2mmol/L dNTP,0.3μmol/L引物,0.5U Taq DNA聚合酶(天根公司),50ng总DNA;扩增程序为94℃ 3min;94℃ 30s,退火温度1min,72℃ 2min,38个循环,72℃ 7min。筛选出了10个适宜山楂遗传分析的ISSR引物。
关键词(KeyWords)： 山楂;ISSR;优化
基金项目(Foundation): 沈阳农业大学青年教师科研基金。
作者(Author): 代红艳;张志宏;周传生;李贺;郭修武;
Email:
DOI:
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