摘要
目的以小麦白粉病抗病品种SARKA和不抗病品种河农822为试验材料,构建定位小麦抗白粉病基因的SSR体系.方法采用单因素筛选的方法,摸索SSR反应体系中各个影响因素的浓度和用量,采用完全随机试验,进行优化组合.结果获得适宜抗白粉病基因定位的SSR标准体系,确定总反应体系为15μL,其中TaqDNA聚合酶0.15μL(5 U/μL),10×PCR Buffer 1.5μL,Mg2+1.08μL(15 mmol/L),引物各0.8μL(25 ng/μL),模板DNA 5μL(10 ng/μL),dNTP 1.2μL(10 mmol/L),ddH2O 4.47μL.结论利用该体系进行扩增,所得谱带清晰、稳定、非特异性带少.
Objective SARKA （Resistance variety） and henong822 （susceptible variety） are used in this study to optimize the SSR Technique System of Powdery Mildew Resistance Genes in Wheat. Methods Single factor selection was made to investigate better concentration and volume of its influencing factors in SSR technique system. Then, thoroughly random experiment was made to seek the best combination a- mong factors. Results At last, the best SSR technique system of resistance genes was established： 0. 15 u/μL （5 U/μL） of Taq DNA polymerase, 1.5 μL10×PCR Buffer, 1.08 μL （15 mmol/L） Mg^2＋, 0.8 μL（25 ng/μL） of each primer, 5 uL （10 ng/μL） of template DNA, 1.2 μL （10 retool/L） of dNTP and 4.47 μL of ddH20 added up to the 15 μL PCR mixture. Conclusion Using this system, amplification bands are clear and stable and non-especial bands are less.
出处
《河北北方学院学报（自然科学版）》
2009年第3期31-35,42,共6页
Journal of Hebei North University:Natural Science Edition
关键词
小麦
白粉病
SSR
体系建立
wheat
powdery mildew
SSR
system establishing
