长果种黄麻DNA的提取及SRAP分子标记体系的建立

Extraction of Genomic DNA from Corohorus olitorius L. and Establishment of SRAP Reaction System

以宽叶长果和甜麻杂交产生的F2代为材料,研究长果种黄麻DNA的提取方法和SRAP分子标记技术中的主要影响因素(包括模板DNA浓度、Mg2+浓度、Taq DNA聚合酶浓度、dNTPs浓度及上下游引物浓度)。最终建立了适于长果种黄麻DNA提取的CTAB法与SRAP-PCR反应体系(25μL):模板DNA 100ng,引物浓度0.48μmol.L-1,Mg2+浓度2.8mmol.L-1,dNTPs浓度0.35mmol.L-1,Taq DNA聚合酶0.7U。

In this study,extract high quality genomic DNA from jute（Corohorus olitorius L） and establish a stable SRAP reaction system,including template DNA concentration,Mg2＋ concentration,Taq DNA polymerase concentration,dNTPs concentration and forward and reverse primer concentration,using a population F2 progeny derived from a cross of glycanes thesia（wild species） and wild leaf jute（cultivated species）.The results showed that the DNA isolated with modified CTAB method had good quality.The optimum SRAP reaction system could amplify high levels of polymorphism,good repeatability and clear band pattern.SRAP-PCR system（total volume of 25 μl） was established as follows： template DNA 100 ng,forward primer 0.48 μmol·L-1,reverse primer 0.48 μmol·L-1,Mg2＋ 2.8 mmol·L-1,dNTPs 0.35 mmol·L-1,Taq DNA polymerase 0.7 U.It showed that the optimized system can be applied in the SRAP analysis for Corohorus olitorius L.,which provided technical support for the genetic linkage map construction in the future.