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[目的]建立黑麦特异性PCR反应优化体系。[方法]以普通小麦“中国春”、S165、黑麦、八倍体小黑麦、六倍体小黑麦为试验材料,研究了模板DNA、引物、dNTPs、Mg2+浓度、TaqDNA聚合酶用量及退火温度对黑麦特异性PCR反应体系的影响。[结果]采用改良的CTABDNA微量提取法可以得到高质量的基因组DNA,满足PCR反应模板的要求,黑麦特异PCR扩增反应体系为:在25μl反应体系中,10×缓冲液,1.5mmol/LMgCl2,200μmol/LdNTP,40ng引物,40～60ng模板DNA,1UTaq酶。[结论]建立了适宜的黑麦特异PCR扩增反应体系,可为小麦背景下黑麦外源种质的检测奠定基础。
[Objective] The research aimed to establish a rye-specific PCR reaction optimization system. [Method] The effects of template DNA, primers, dNTPs, Mg2 + concentration, Taq DNA polymerase Effect of dosage and annealing temperature on rye - specific PCR reaction system. [Result] High quality genomic DNA could be obtained by modified CTAB DNA micro-extraction method to meet the requirements of PCR reaction template. The specific rye reaction system was as follows: 10 × buffer, 1.5 mmol / L MgCl 2 , 200μmol / LdNTP, 40ng primer, 40 ~ 60ng template DNA, 1UTaq enzyme. [Conclusion] The suitable rye specific PCR amplification reaction system was established, which could lay the foundation for the detection of exogenous germplasm of rye under the wheat background.