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SSR技术鉴定玉米品种真实性反应体系的优化研究 被引量:6

Optimization of SSR Reaction System for Identifying the Authenticity of Maize Hybrids
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摘要 [目的]优化SSR技术鉴定玉米品种真实性反应体系。[方法]对SSR技术参数(PCR反应体系、退火温度和电泳时间)进行了优化,并对辽宁省主推的10个玉米品种进行了鉴定。[结果]最优PCR反应体系为:灭菌超纯水14.60μl,10×Buffer(Mg2+)2.00μl,dNTPs 1.20μl,Taq酶0.20μl,正、反向引物各0.50μl及DNA原液1.00μl。退火温度和电泳时间对PCR扩增结果的影响较大,所选的引物不同,在同一条件下呈现理想电泳条带所需的最佳退火温度和电泳时间不同。[结论]该体系应用于杂交玉米品种的真实性快速鉴定是可行的。 [Objective] The aim was to optimize SSR reaction system applied in the identification of authenticity of maize variety.[Method] The technical parameters of SSR including PCR reaction system,annealing temperature and electrophoresis time were optimized to identify 10 major maize varieties in Liaoning Province.[Result] The optimum PCR reaction system was: 14.60 μl sterile ultrapure water,2.00 μl 10 × Buffer(Mg2+),1.20 μl dNTPs,0.20 μl Taq enzyme,0.50 μl each of the forward and reverse primers and 1.00 μl DNA stock solution.Annealing temperature and electrophoresis time could greatly influenced the results of PCR amplification.The optimal annealing temperature and electrophoresis time required for the ideal electrophoresis bands under the same conditions were different when different primers were used.[Conclusion] The system is feasible to apply in rapid identification of authenticity of hybrid maize varieties.
出处 《安徽农业科学》 CAS 北大核心 2011年第9期5111-5113,共3页 Journal of Anhui Agricultural Sciences
关键词 玉米 SSR标记 真实性 优化 Maize(Zea mays L.) Simple sequence repeat Marker Authenticity Optimization
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  • 1SENIOR M L,MURPHY J P,GOODMAN M M,et al.Utility of SSRs for determining genetic similarities and relationships in maize using an agarose gel system[J].Crop Sci,1998,38:1088-1098.
  • 2袁力行,傅骏骅,Warburton M,李新海,张世煌,Khairallah M,刘新芝,彭泽斌,李连城.利用 RFLP、SSR、AFLP和 RAPD标记分析玉米自交系遗传多样性的比较研究[J].Acta Genetica Sinica,2000,27(8):725-733. 被引量:259
  • 3JINNEMAN K C,YOSHTTOMI K J,WEAGANT S D.Multiplex real time PCR method toidemify shiga toxin genes stxl and stx2 and Escherichia coli 0157:H7/H-serotype[J].Applied and EnvironmentalMicrobiology,2003,69(10):6327-6333.

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