摘要
采用单因素试验和正交设计相结合进行了侧耳(Pleurotus ostreatus)RSAP-PCR反应体系的建立和优化,并对优化后体系的稳定性进行了验证。讨论了Mg2+浓度、dNTPs浓度、引物浓度、Taq聚合酶用量和模板DNA含量对侧耳RSAP-PCR反应的影响,最终得出总体积为25μL的反应体系:30 ng模板DNA,2.0μL10倍PCR buffer,Mg2+2.5 mmol/L,dNTPs 0.25 mmol/L,引物0.32μmol/L,Taq DNA聚合酶2U,ddH2O 13.2μL。
In this study, single factor and orthogonal experiments were used to establish and optimize the RSAP- PCR reaction system of Pleurotus ostreatus. Then, experiment tests were done to check the sta- bility of the RSAP - PCR reaction system using a different pair of primers. Finally, a 25μL optimized re- action system was obtained： 30 ng DNA, 2.0 μL 10 × PCR buffer, Mg2＋ 2.5 mmol/L, dNTPs 0.25 mmol/L, Primers 0.32 μmol/L, Taq DNA polymerase 2 U, ddH20 13.2μL.
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2013年第2期171-176,共6页
Journal of Jilin Agricultural University
基金
吉林省农委项目(10096)
关键词
侧耳
RSAP
体系优化
Pleurotus ostreatus
RSAP
system optimization
