title: Cloning and  sequencing of the rDNA gene family of the water buffalo (Bubalus bubalis).
abstract: 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.
authors.affiliation: Guangxi Key Laboratory of Buffalo Genetics, Breeding and Reproduction, Guangxi Buffalo Research Institute, Chinese Academy of Agricultural Science, Nanning, P.R. China.
authors.forename: C Y
authors.initials: CY
authors.lastname: Pang
authors.forename: T X
authors.initials: TX
authors.lastname: Deng
authors.forename: D S
authors.initials: DS
authors.lastname: Tang
authors.forename: C Y
authors.initials: CY
authors.lastname: Yang
authors.forename: H
authors.initials: H
authors.lastname: Jiang
authors.forename: B Z
authors.initials: BZ
authors.lastname: Yang
authors.forename: X W
authors.initials: XW
authors.lastname: Liang
mesh_terms: D000818:Animals; D002020:Buffaloes / Q000235:genetics*; D003001:Cloning, Molecular; D004275:DNA, Ribosomal / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D004996:Ethidium; D019143:Evolution, Molecular; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics*; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D017422:Sequence Analysis, DNA / Q000379:methods*; D014158:Transcription, Genetic
chemical_list: D004275:DNA, Ribosomal; D012337:RNA, Ribosomal, 18S; D004996:Ethidium