摘要
以油茶品种大别山2号和赣兴46为试验材料,利用Me5Em8正反向引物组合进行了SRAP-PCR反应体系的L9(34)正交试验,采用正交设计直观分析及方差分析对影响SRAP反应的Taq聚合酶、Mg2+、dNTP和引物浓度进行分析,并对模板DNA浓度进行了单因素试验分析。结果表明,油茶SRAP-PCR 20μL反应体系的最佳组合为:Taq聚合酶1.20μmol/min、Mg2+浓度1.25 mmol/L、dNTP浓度0.15 mmol/L、Primer浓度0.60μmol/L、模板DNA含量60 ng,并含有2μL 10×buffer(Mg2+free)。各因素对油茶SRAP-PCR反应的影响大小依次为:dNTP、Mg2+、Taq聚合酶、Primer。
In this paper,orthogonal design to SRAP-PCR system for Camellia oleifera cultivar Dabieshan 2 and Ganxing 46 was conducted with the primers combination of Me5 and Em8.Orthogonal design-direct analysis and variance analysis were applied to optimize SRAP-PCR amplification system in 4 factors such as Taq DNA polymerase,Mg2＋,dNTP and primer.Template DNA was also analyzed by the single element test.The results indicated that an optimal reaction system（20 μL）of SRAP-PCR system was established： 1.20 μmol/min Taq DNA polymerase 1.25 mmol/L Mg2＋,0.15 mmol/L dNTP,0.60 μmol/L primer,60 ng template DNA and 2 μL 10 buffer（Mg2＋ free）.And,the order of each factor levels affected on the result of SRAP-PCR was dNTP,Mg2＋,Taq DNA polymerase and primer.
出处
《南京林业大学学报（自然科学版）》
CAS
CSCD
北大核心
2011年第5期112-116,共5页
Journal of Nanjing Forestry University：Natural Sciences Edition
基金
"十一五"国家科技支撑计划(2009BADB1B04)
关键词
油茶
SRAP
反应体系
正交设计
Camellia oleifera
SRAP
reaction system
orthogonal design
