沙棘SRAP-PCR反应体系的优化及引物筛选

摘要

The concentrations of Mg2+, Taq DNA polymerase, primers, dNTPs, template DNA affecting the SRAP-PCR reaction were optimized to establish the SRAP-PCR system for sea buckthorn (Hippophae rhamnoides L). The optimum 25 μL reaction system contained 3.0 mmol/L Mg2+, 2 U/25 ΜL Taq DNA polymerase, 0.8 μmol/L primers, 0.25 mmol/L dNTPs and 30 ng/25 μL DNA templates. 17 out of 30 primer pair combinations were screened out using the optimized amplification system, which would support the molecular marker assisted breeding of sea buckthorn.%为建立适合沙棘(Hippophae rhamnoides L.)的SRAP-PCR反应体系,对影响SRAP-PCR的Mg2+浓度、Taq DNA聚合酶浓度、引物浓度、dNTPs浓度、模板DNA浓度进行了优化.优化后的反应体系为Mg2+3.0 mmol/L、Taq DNA聚合酶2U/25μL、引物0.8 μmol/L、dNTPs 0.25 mmol/L、模板DNA 30ng/25μL,反应总体系25 μL.利用此体系从30对引物组合中筛选出17对适合沙棘SRAP-PCR的引物,有助于沙棘的分子标记辅助育种研究.