摘要
为了获得最佳的克氏原螯虾ISSR-PCR反应体系,采用单因素实验法将反应体系的Taq聚合酶、dNTPs和Mg2+3个主要因素设定5个梯度,根据每个因素量的变化对扩增结果产生的影响进行了研究,最后确定最佳反应体系为:总体积10μL,1μL10×TaqBuffer、dNTPs浓度0.6mmol/L、0.6U Taq DNA polymerase、MgCl2浓度1.8mmol/L、20ngDNA,引物浓度0.8μmol/L。
For optimizing ISSR-PCR reaction system of Procambarus clarkii,the single factor experiment included three factors,such as Taq DNA polymerse,dNTPs and Mg 2＋,to five different concentrations were set to study the effect of these variations in ISSR-PCR amplification in this research.In conclusion the reaction system was optimized as below：1 μL 10 × Taq Buffer,0.6 mmol/L dNTPs,0.6 U Taq DNA polymerase,1.8 mmol/L MgCl 2,20 ng DNA and 0.8 μmol/L primer in 10 μL reaction solution.
出处
《中国农学通报》
CSCD
北大核心
2010年第21期432-435,共4页
Chinese Agricultural Science Bulletin
基金
国家科技支撑计划资助项目"长江中游地区特色水产品高效生产关键技术研究与示范"之课题"克氏原螯虾高效无公害生产关键技术研究与示范"(2007BAD37B01)
