摘要
[Objective] This study aimed optimize sequence-related amplified polymor- phism （SRAP）-PCR system for Paphiopedilum hirsutissimum. [Method] Using P. hir- sutissimum leaf as the material, a single-factor test was adopted to optimize the fac- tors in SRAP-PCR system, including the concentrations of dNTPs, Mg2＋, Taq poly- merase, DNA template and primers. [Result] The optimized SRAP-PCR system con- tained 2.5 μl of 10xPCR buffer, 0.15 mmol/L dNTPs, 2.0 mmol/L Mg2＋, 0.3 mmol/L primer each, 0.3 U Taq polymerase and 1 μl of DNA template. [Conclusion] This system can amplify clear and repeatable DNA profiles, which can be applied for further study about P. hirsutissimum.
[目的]确定带叶兜兰的SRAP反应体系。[方法]以带叶兜兰嫩叶提取的DNA为材料,对带叶兜兰SRAP反应体系中的主要成分dNTPs、Mg2+、Taq酶、模板DNA及引物进行了单因子优化。[结果]最终确定了适合带叶兜兰基因扩增的SRAP反应体系:在25μl反应体系中加入10×PCR Buffer 2.5μl、dNTPs0.15 mmol/L、Mg2+2.0 mmol/L、引物0.4 mmol/L、Taq酶0.3 U。[结论]该体系扩增条带清晰,重复性好,可应用于带叶兜兰的多方面的分析。
基金
Supported by Scientific Research Fund for the Introduced Talents in Hechi University(2008QB-N001)~~
