海南普通野生稻ISSR-PCR反应体系的优化  

STUDY ON OPTIMIZATION FOR ISSR-PCR REACTION SYSTEM FOR WILD RICE IN HAINAN PROVINCE

作　　者：韩飞[1,2] 侯立恒[2] 李敬娜[2] 袁潜华[1,2] 

机构地区：[1]海南大学热带生物资源教育部重点实验室,海南海口570228 [2]海南大学农学院,海南海口570228

出　　处：《贵州科学》2008年第4期40-44,共5页Guizhou Science

基　　金：国家863计划项目(2002AA212091)

摘　　要：通过不同浓度的Mg2+、模板DNA、dNTPs、Taq酶、引物、退火温度和循环次数的单因素试验,筛选海南普通野生稻最佳ISSR-PCR反应体系为:15μl反应体系中含2.0 mmol/L MgCl2,50 ng模板DNA,200μmol/L dNTPs,1.20 U Taq酶,1.5μmol/L ISSR引物,50 mmol/L KCl 10 mmol/L Tris-HCl(pH8.3).反应程序:94℃预变性5 min;94℃,1 min,52℃,1 min,72℃,2 min,35个循环;72℃延伸10min,4℃保存.To optimize ISSR-PCR reaction system for wild rice in Hainan Province,the effects of seven factors Mg2+,dNTPs,Taq DNA polymerse,primer,T-DNA,annealing temperature and cycle were studied at six levels respectively.The results showed that optimum ISSR-PCR reaction system of wild rice was every 15 μL reaction system containing 2.0 mmol/L MgCl2,50ng template DNA,200μmol/L dNTPs,1.20U Taq DNA polymerse,1.5 μmol/L ISSR primer,50 mmol/L KCl,10 mmol/L Tris-HCl(pH 8.3).The above reaction mixture was denatured at 94℃...

关 键 词：海南 普通野生稻 ISSR 反应体系 PCR 

分 类 号：S511.9[农业科学—作物学]