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食品研究与开发:2018,39(2):159-163
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实时荧光定量PCR检测产琥珀酸丝状杆菌数量方法的研究
王国泽1,2,刘达玉2,余华2,郭秀兰2,左福元1,*,朱智1
(1.西南大学,重庆402460;2.成都大学药学与生物工程学院,四川成都610106)
Studies on Using Quantitative Real-time PCR to Detect the Amount of Fibrobacter succinogenes
WANG Guo-ze1,2,LIU Da-yu2,YU Hua2,GUO Xiu-lan2,ZUO Fu-yuan1,*,ZHU Zhi1
(1.Southwest University,Chongqing 402460,China;2.College of Pharmacy and Biological Engineering,Chengdu University,Chengdu 610106,Sichuan,China)
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投稿时间:2017-05-09    
中文摘要: 为建立产琥珀酸丝状杆菌定性定量研究方法,实现后期该菌种用于纤维素生物可再生能源等方面应用研究。本试验提取15份瘤胃细菌脱氧核糖核酸(Deoxyribonucleic acid,DNA),利用SYBR Premix Ex Taq荧光染料嵌合法和人工合成基因技术,采用实时荧光定量PCR方法检测样品中产琥珀酸丝状杆菌数量;结果:所构建的质粒DNA标准曲线线性关系良好,R2=1,所有样品溶解曲线均为单峰,扩增曲线呈S型,扩增效率为93.9%。表明应用构建实时荧光定量PCR法可对实际样品中产琥珀酸丝状杆菌进行定性定量研究。
Abstract:For building the qualitative and quantitative analysis of Fibrobacter succinogenes,implement the bacteria used on the cellose biological renewable energy late application research.This experiment extracted 15 rumen bacterias'DNA,used SYBR Premix Ex Taq fluorescent legal and synthetic gene technology,embedded quantitati-ve Real-time PCR to detect the amounts of Fibrobacter succinogenes in samples.Rested:the constructed plasmid DNA standard curve linear relationship was good,R2=1,all samples'dissolution curves were unimodal and S-shaped curve types,amplification efficiency was 93.9%.The study that built of quantitative real-time PCR was Indicated to be able to the qualitative and quantitative analysis of Fibrobacter succinogenes in the actual samples.
文章编号:201802028     中图分类号:    文献标志码:
基金项目:重庆市高校创新团队资助(CXTDG201602003);四川省教育厅一般项目(荧光定量PCR检测瘤胃内纤维可再生能源降解菌体系研究)
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