杏ISSR反应体系的优化和指纹图谱的构建Optimization of ISSR reaction system and construction of cultivar fingerprint in apricot
刘威生,冯晨静,杨建民,刘冬成,张爱民,李绍华
摘要(Abstract):
以杏品种清密沙为试材,用引物UBC825〔序列为(AC)8T〕研究了PCR反应体系的主要成分及退火温度对杏ISSR扩增结果的影响。结果表明:Primer、Mg2+、Taq酶浓度对扩增效果有明显影响,而模板DNA和dNTPs含量对扩增结果影响不大。优化的反应体系为:20μL的反应体系中含10ng模板DNA、0.1mmol/LdNTP、0.25μmol/LPrimer、2.5mmol/LMg2+,0.5UTaqPolymerase。适宜退火温度为50-52.1℃。用引物UBC825和UBC868〔序列为(GAA)5〕在优化的反应条件下建立了5个种12份杏材料的ISSR指纹图谱。
关键词(KeyWords): 杏;ISSR(inter-simple sequence repeat);反应体系;退火温度;优化
基金项目(Foundation):
作者(Author): 刘威生,冯晨静,杨建民,刘冬成,张爱民,李绍华
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参考文献(References):
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