Cloning and sequencing of the rDNA gene family of the water buffalo (Bubalus bubalis)

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 rD...

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Bibliographic Details
Published in:Genetics and molecular research 2012-01, Vol.11 (3), p.2878-2883
Main Authors: Pang, C Y, Deng, T X, Tang, D S, Yang, C Y, Jiang, H, Yang, B Z, Liang, X W
Format: Article
Language:English
Subjects:
Animals
Buffaloes - genetics
Cloning, Molecular
DNA, Ribosomal - genetics
Electrophoresis, Agar Gel
Ethidium
Evolution, Molecular
Molecular Sequence Data
Multigene Family - genetics
Phylogeny
Polymerase Chain Reaction
Repetitive Sequences, Nucleic Acid
RNA, Ribosomal, 18S - genetics
Sequence Analysis, DNA - methods
Transcription, Genetic
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Summary: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.
ISSN:1676-5680
1676-5680
DOI:10.4238/2012.July.10.3