摘要
以冬枣DNA为研究对象,利用单因素试验,分析了Mg2+浓度、dNTP浓度、Taq酶含量、引物浓度、模板DNA含量以及退火温度对ISSR-PCR扩增的影响,经优化建立了枣属植物最适ISSR-PCR反应体系,25μL反应液中包含1×PCR Buffer、2.0 mmol/L Mg2+、0.25 mmol/L dNTP、1.5 UTaq酶、1.25μmol/L引物和50 ng模板,最适退火温度为58℃。PCR反应体系的建立为应用ISSR标记技术开展枣种群遗传变异分析和构建遗传图谱奠定基础。
Take the genomic DNA of Ziziphus jujuba Mill.cv.Dongzao as a template,the ISSR-PCR reaction system of Chinese jujube was estabblished after analysised the effect of 6 main factors in PCR reaction using the single factor method.The optimal ISSR-PCR reaction system was determined as follows：1×PCR Buffer,2.0 mmol/L Mg2＋,0.25 mmol/L dNTP,1.5 U Taq polymerase,1.25 μmol/L primer and 50 ng template DNA in total 25 μL reaction volume.The optimal annealing temperature was 58℃.All the result established a foundation for building cultivar fingerprinting and genetic linkage map in Chinese jujube by using ISSR molecular markers.
出处
《华北农学报》
CSCD
北大核心
2008年第B10期209-212,共4页
Acta Agriculturae Boreali-Sinica
基金
国家自然科学基金项目(30370994)
关键词
枣
ISSR
体系优化
Chinese jujube
ISSR
System optimization
