变性梯度凝胶电泳(Denaturing gradient gel electrophoresis,DGGE)技术是最先用于DNA突变检测的一种电泳技术。为获得最佳的PCR-DGGE上样模板,本实验分别采用十六烷基三甲基溴化铵(CTAB)法和Power Soil DNA Isolation Kit试剂盒抽提细...变性梯度凝胶电泳(Denaturing gradient gel electrophoresis,DGGE)技术是最先用于DNA突变检测的一种电泳技术。为获得最佳的PCR-DGGE上样模板,本实验分别采用十六烷基三甲基溴化铵(CTAB)法和Power Soil DNA Isolation Kit试剂盒抽提细菌基因组DNA,分别以Primer(338F/518R)/Primer(P2/P3)为16Sr DNA V3扩增引物,采用温度梯度PCR对引物扩增条件进行优化。结果显示:在Tm 56℃,通过CTAB法抽提细菌基因组DNA,采用primer(338F/518R)扩增16Sr DNA V3目的产物特异性最佳。展开更多
Twenty-nine facultative anaerobic cellulose-degrading bacteria were isolated from soil, rumen fluid, rumen residues and diet of dairy cow. Based on 16 Sr DNA analysis by BLAST algorithm method, the results showed that...Twenty-nine facultative anaerobic cellulose-degrading bacteria were isolated from soil, rumen fluid, rumen residues and diet of dairy cow. Based on 16 Sr DNA analysis by BLAST algorithm method, the results showed that most of the strains were Bacillus genera, and six of the 29 strains were bigger than 10 mm of diameter of clear zones. For them, two strains were isolated from rumen fluid(L5 and L7) and other two were isolated from rumen residue(N5 and N9), while others were isolated from soil(T1) and diet(S6), respectively. Strains from rumen fluid and residue had higher activities of FPCase and CMCase, but lower β-glucosidases.展开更多
文摘变性梯度凝胶电泳(Denaturing gradient gel electrophoresis,DGGE)技术是最先用于DNA突变检测的一种电泳技术。为获得最佳的PCR-DGGE上样模板,本实验分别采用十六烷基三甲基溴化铵(CTAB)法和Power Soil DNA Isolation Kit试剂盒抽提细菌基因组DNA,分别以Primer(338F/518R)/Primer(P2/P3)为16Sr DNA V3扩增引物,采用温度梯度PCR对引物扩增条件进行优化。结果显示:在Tm 56℃,通过CTAB法抽提细菌基因组DNA,采用primer(338F/518R)扩增16Sr DNA V3目的产物特异性最佳。
基金Supported by Northeast Agricultural University Innovation Funding for Postgraduate(yjscx14007)China Agriculture Research System(CARS-37)
文摘Twenty-nine facultative anaerobic cellulose-degrading bacteria were isolated from soil, rumen fluid, rumen residues and diet of dairy cow. Based on 16 Sr DNA analysis by BLAST algorithm method, the results showed that most of the strains were Bacillus genera, and six of the 29 strains were bigger than 10 mm of diameter of clear zones. For them, two strains were isolated from rumen fluid(L5 and L7) and other two were isolated from rumen residue(N5 and N9), while others were isolated from soil(T1) and diet(S6), respectively. Strains from rumen fluid and residue had higher activities of FPCase and CMCase, but lower β-glucosidases.
