安徽农业科学2013，Vol.41Issue(6)：2471-2473,3.
琥珀酸丝状杆菌RT-PCR方法的建立
Development of a RT-PCR Approach for Quantification of Fibrobacter succinogene
摘要
Abstract
[Objective] The research aimed to study the changes of microbe in cattle rumen. [ Method ] Using Fibrobacter succinogene as the research object, a pair of primers was designed based on 16S rRNA gene sequences of F. succinogene. Total DNA was extracted from cattle rumen liquid to amplify DNA fragments of F. succinogene and connected with pDM-18T vector. After PCR and sequencing, recombinant plas-mids with different dilution degree was used as templates to make RT- PCR. [ Result ] The homology between the amplified target fragment and the related fragment of the known strain was 100% . The amplification curves of RT- PCR established with recombinant pMD-18T with different dilution degree as templates had obvious differences. The correlation coefficient of the standard curve was close to 1. The melting curves of RT-PCR displayed a single peak. [Conclusion] The real-time fluorescence quantitative PCR was successfully established for F. succinogene, which laid foundation for further study on the changes of microbe in rumen.关键词
琥珀酸丝状杆菌/RT-PCR/标准曲线Key words
Fibrobacter succinogene/RT-PCR/Standard curve分类
农业科技引用本文复制引用
周焱伟,贺丛,许利凡,王绿林,吴胜强,邓立新,王世根..琥珀酸丝状杆菌RT-PCR方法的建立[J].安徽农业科学,2013,41(6):2471-2473,3.基金项目
现代农业(肉牛牦牛)产业技术体系建设专项(CARS-38). （肉牛牦牛）
