农业科学与技术（英文版）
農業科學與技術（英文版）
농업과학여기술（영문판）
AGRICULTURAL SCIENCE & TECHNOLOGY
2011年
2期
201-204
,共4页
黑麦%PCR反应体系%优化
黑麥%PCR反應體繫%優化
흑맥%PCR반응체계%우화
Rye%PCR reaction system%Optimization
[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CTAB DNA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,1.5 mmol/L MgCl2,200μmol/L dNTP,40 ng引物,40～60ng模板DNA,1 U Taq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.
[目的]建立黑麥特異性PCR反應優化體繫.[方法]以普通小麥"中國春"、S165、黑麥、八倍體小黑麥、六倍體小黑麥為試驗材料,研究瞭模闆DNA、引物、dNTPs、Mg2+濃度、TaqDNA聚閤酶用量及退火溫度對黑麥特異性PCR反應體繫的影響.[結果]採用改良的CTAB DNA微量提取法可以得到高質量的基因組DNA,滿足PCR反應模闆的要求,黑麥特異PCR擴增反應體繫為:在25μl反應體繫中,10×緩遲液,1.5 mmol/L MgCl2,200μmol/L dNTP,40 ng引物,40～60ng模闆DNA,1 U Taq酶.[結論]建立瞭適宜的黑麥特異PCR擴增反應體繫,可為小麥揹景下黑麥外源種質的檢測奠定基礎.
[목적]건립흑맥특이성PCR반응우화체계.[방법]이보통소맥"중국춘"、S165、흑맥、팔배체소흑맥、륙배체소흑맥위시험재료,연구료모판DNA、인물、dNTPs、Mg2+농도、TaqDNA취합매용량급퇴화온도대흑맥특이성PCR반응체계적영향.[결과]채용개량적CTAB DNA미량제취법가이득도고질량적기인조DNA,만족PCR반응모판적요구,흑맥특이PCR확증반응체계위:재25μl반응체계중,10×완충액,1.5 mmol/L MgCl2,200μmol/L dNTP,40 ng인물,40～60ng모판DNA,1 U Taq매.[결론]건립료괄의적흑맥특이PCR확증반응체계,가위소맥배경하흑맥외원충질적검측전정기출.
[Objective] The aim was to establish the optimal rye-specific PCR reaction system for rye. [Method] The ordinary wheat "Chinese Spring", S165, rye, octoploid triticale and hexaploid triticale were used as materials to carry out study on the effect of the amount of template DNA, primers, dNTPs, Mg2+ concentrations, Taq DNA polymerase and annealing temperature on the rye-specific PCR reaction system of rye.[Result] The genomic DNA extracted by modified CTAB DNA extraction method showed high quality, which was satisfied for the PCR reaction template. The rye-specific PCR reaction system was 25 μl, including 10 × buffer solution, 1.5 mmol/L MgCI2, 200 μmol/L dNTP, 40 ng primers, 40-60 ng DNA template and 1 U Taq DNA polymerase. [Conclusion] The optimal rye-specific PCR reaction system was established,which provided basis for the identification of exogenous germplasm of rye in wheat background.
