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洋葱Ms位点多重PCR标记的开发与优化
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作者 王振宝 霍雨猛 +7 位作者 刘冰江 缪军 杨妍妍 高莉敏 孔素萍 程斐 陈宁 吴雄 《山东农业科学》 2014年第9期7-11,共5页
以本课题组已获得的洋葱Ms位点侧翼序列为基础,设计、筛选引物获得了一个兼容的多重PCR分子标记,并对其反应体系和反应程序进行了优化。优化后的扩增体系:10×PCR buffer(Mg2+free)2.5μL、25 mmol/L MgCl24μL、2.5 mmol/L dNTP 6... 以本课题组已获得的洋葱Ms位点侧翼序列为基础,设计、筛选引物获得了一个兼容的多重PCR分子标记,并对其反应体系和反应程序进行了优化。优化后的扩增体系:10×PCR buffer(Mg2+free)2.5μL、25 mmol/L MgCl24μL、2.5 mmol/L dNTP 6μL、DNA模板1μL(约50 ng)、10μmol/L引物各1μL、5 U/μL rTaq聚合酶0.6μL、用灭菌双蒸水补齐至25μL;反应程序:94℃预变性5 min;94℃变性30 s,65.4℃退火45 s,72℃延伸1 min,35个循环;最后72℃延伸10 min。优化后的体系和程序可以检测到清晰的目的条带,通过一次PCR反应即可鉴定Ms位点的3种基因型(MsMs、Msms、msms),操作简单,稳定性好。 展开更多
关键词 洋葱 雄性不育 Ms位点 多重pcr标记
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Establishment and Application of a Multiplex PCR System for the Detection of Blast Resistance Genes Pi-ta and Pi-b in Rice 被引量:1
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作者 姚姝 刘燕清 +7 位作者 张亚东 朱镇 陈涛 赵庆勇 周丽慧 赵春芳 于新 王才林 《Agricultural Science & Technology》 CAS 2015年第10期2225-2230,共6页
Rice blast is one of the important diseases in major rice producing areas of China. The main blast resistance genes Pi-ta and Pi-b showed broad-spectrum and durable resistance to rice blast in many rice growing areas ... Rice blast is one of the important diseases in major rice producing areas of China. The main blast resistance genes Pi-ta and Pi-b showed broad-spectrum and durable resistance to rice blast in many rice growing areas of China, which have been widely utilized in rice breeding and commercial production. In this study, on the basis of detection and verification of the genotypes of 22 rice varieties har- boring known blast resistance genes (Pi-ta and Pi-b) and blast susceptibility genes (pi-ta and pi-b), two multiple PCR systems for these genes were established by us- ing the functional markers of blast resistance genes Pi-ta and Pi-b as well as blast susceptibility genes pi-ta and pi-b, respectively. Specifically, multiple PCR system I could simultaneously detect blast resistance genes Pi-ta and Pi-b, while system II could detect simultaneously blast susceptibility genes pi-ta and pi-b. In addition, the genotypes of 336 high generation breeding materials were detected with these two multiple PCR systems. The results were highly consistent with those of conventional single mark detection, indicating that these two multiplex PCR systems were stable, reliable and time-saving. The established multiplex PCR systems may serve as a rapid and efficient method to identify and screen rice germplasm resources and can be applied in marker-assisted selection to polymerize multiple genes for blast resis- tance in rice breeding. 展开更多
关键词 Rice blast Molecular marker Resistance gene Multiplex pcr system
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