文献类型： 外文期刊
作者： Pang, C. Y. 1 ; Deng, T. X. 1 ; Tang, D. S. 2 ; Yang, C. Y. 1 ; Jiang, H. 2 ; Yang, B. Z. 1 ; Liang, X. W. 1 ;
作者机构： 1.Chinese Acad Agr Sci, Guangxi Buffalo Res Inst, Guangxi Key Lab Buffalo Genet Breeding & Reprod, Nanning, Peoples R China
2.Foshan Univ, Coll Med, Foshan, Peoples R China
关键词： Molecular cloning;GC-rich;rDNA;rRNA;Bubalus bubalis
期刊名称：GENETICS AND MOLECULAR RESEARCH （ 影响因子：0.764； 五年影响因子：0.912 ）
ISSN： 1676-5680
年卷期： 2012 年 11 卷 3 期
页码：
收录情况： SCI
摘要： The rDNA genes coding for ribosomal RNA in animals are complicated repeat sequences with high GC content. We amplified water buffalo rDNA gene sequences with the long and accurate (LA) PCR method, using LA Taq DNA polymerase and GC buffer, based on bioinformatic analysis of related organisms. The rDNA genes were found to consist of 9016 nucleotides, including three rRNA genes and two internal transcribed spacers (ITS), which we named 18S rRNA, ITS1, 5.8S rRNA, ITS2 and 28S rRNA. We tested and optimized conditions for cloning these complicated rDNA sequences, including specific rules of primer design, improvements in the reaction system, and selection of the DNA polymerase.
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