期刊论文

Establishment and optimization of SRAP reaction system in Castanea henryi

中文

经济林研究

1003-8981

2014

3

8-15

10.14067/j.cnki.1003-8981.2014.03.039

国家“十二五”科技支撑计划课题“板栗与锥栗高效生产关键技术研究与示范”（2013BAD14B04）；

本校为第一机构

为了建立锥栗SRAP-PCR反应体系,从而为今后的锥栗分子生物学研究提供理论参考,以锥栗叶片基因组DNA为材料,采用单因素试验和正交试验相结合的研究方法,对影响锥栗SRAP-PCR反应体系的模板用量、Mg^2＋浓度、dNTP浓度、引物浓度和Taq DNA聚合酶浓度这5个主要因素进行了优化试验。试验确立的适合锥栗的SRAP-PCR反应体系为：20μL体系中,模板DNA用量为40 ng,Mg^2＋浓度为1.8 mmol/L,dNTP浓度为280μmol/L,Taq DNA聚合酶浓度为2.0 U,引物浓度为0.6μmol/L。

In order to establish a SRAP-PCR reaction system, and provide theoretical references for the molecular biology research on Castanea henryi in future, taking the genomic DNA of C. henryi leaves as material, the fi ve main factors of concentrations of template DNA, Mg^2＋, dNTP, primer and Taq DNA polymerase in SRAP-PCR amplifi cation system were optimized by using the single factor orthogonal design. The SRAP-PCR amplifi cation conditions suitable for C. henryiare established, containing 40 ng template DNA, 1.8 mmol/L Mg^2＋, 280 μmol/L dNTP, 2.0...