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[目的]筛选经济、稳定性好的水稻PCR反应体系,并检测所选体系在不同的基于PCR反应的分子标记中的通用性。[方法]以CTAB法提取的水稻叶片DNA为模板,应用L16(45)正交设计进行PCR反应体系优化。[结果]16个不同处理组合均扩增出了清晰谱带,但扩增效果及PCR产量有差异。最经济、适用的体系:20μl反应体系,20ng模板DNA,浓度150μmol/LdNTP,浓度0.2μmol/L引物,1.0UTaqDNA聚合酶,浓度1.5mmol/LMg2+,1×Taqbuffer,剩余体积用超纯水补齐。[结论]试验确定了水稻PCR优化反应体系,该体系也适用于一些其他基于PCR反应的标记。
[Objective] The research aimed to screen economical and stable rice PCR reaction system and to test the versatility of the selected system in different PCR-based molecular markers. [Method] The DNA of rice leaves extracted by CTAB method was used as template and the PCR reaction system was optimized by L16 (45) orthogonal design. [Result] The clear bands were amplified in all 16 different combinations, but the amplification effect and PCR yield were different. The most economical and suitable system: 20μl reaction system, 20ng template DNA, concentration 150μmol / LdNTP, 0.2μmol / L primer, 1.0UTaq DNA polymerase, concentration 1.5mmol / LMg2 +, 1 × Taqbuffer, the remaining volume is filled with ultrapure water . [Conclusion] The experiment determined rice PCR optimization reaction system, and the system was also suitable for some other markers based on PCR reaction.