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Genetic Characterization Of Pakistani Buffalo Breeds By Mitochondrial D-Loop And Microsatellite Analyses

By: Tanveer Hussain | Prof.Dr.Masroor Elahi Babar.
Contributor(s): Dr. Khalid Javed | Prof. Dr. Irshad Hussain.
Material type: materialTypeLabelBookPublisher: 2008Subject(s): Department of Livestock ProductionDDC classification: 1114,T Dissertation note: Pakistan has various dairy breeds of buffalo and cattle, but the genetic data of different buffalo breeds like Nih, Ravi, Nihi-Ravi, Kundi and Azakheli is lacking which need to be established for their genetic characterization. Blood samples of unrelated true representatives of all breeds were collected from their respective home tracts i.e Nih Ravi (LPRI Bahadarnagar, Okara, BRI Pattoki, Rakh Dera Chahi, Lahore); Nih (Pakpatan, Minchnabad, Arifwala, Hasilpur); Ravi (Kamahia, Tandlianwala); Kundi (Tandojam, Tando Muhammad Khan, Dadu) and Azakheli (Directorate of Livestock Research & Development Station Surezai, Peshawar and Matta, Swat). DNA was extracted with the use of standard protocol and amplification of the mitochondrial D-loop region was done with specific primers in Molecular Cytogenetics and Genomics Laboratory in the department of Livestock Production. Sequencing of amplified portion of mt DNA D-loop was done. Sequences were analyzed with the help of software blast2sequence. Single Nucleotide Polymorphisms (SNPs) were identified and comparison of 52 mitochondrial DNA haplotypes of all buffalo breeds was done. Genetic distance and identity between five buffalo breeds were calculated and phylogenetic tree was constructed using BioEdit and MEGA 4.1 softwares showing the relationships between different haplotypes. Domestication events were also observed through network analysis. For further confirmation of the genetic structure of buffalo breeds 8 dye labeled microsatehhite markers (recommended by ISAG) were used and genotyping was done. Results were analyzed with the help of different softwares. Genetic diversity, Allele frequencies, observed and expected homozygosity and heterozygosity, Hardy Weinberg equilibrium, F-Statistics and Gene Flow for all Loci, population's dendogram, Neis genetic identity and genetic distance/ diversity was calculated. This work provided the genetic data which is very helpful for determining the genetic diversity of buffalo population, breed identification, animal forensic and paternity cases and making effective breeding policies and conservational activities in future.
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Pakistan has various dairy breeds of buffalo and cattle, but the genetic data of different buffalo breeds like Nih, Ravi, Nihi-Ravi, Kundi and Azakheli is lacking which need to be established for their genetic characterization. Blood samples of unrelated true representatives of all breeds were collected from their respective home tracts i.e Nih Ravi (LPRI Bahadarnagar, Okara, BRI Pattoki, Rakh Dera Chahi, Lahore); Nih (Pakpatan,

Minchnabad, Arifwala, Hasilpur); Ravi (Kamahia, Tandlianwala); Kundi (Tandojam, Tando Muhammad Khan, Dadu) and Azakheli (Directorate of Livestock Research & Development Station Surezai, Peshawar and Matta, Swat). DNA was extracted with the use of standard protocol and amplification of the mitochondrial D-loop region was done with specific primers in Molecular Cytogenetics and Genomics Laboratory in the department of Livestock Production. Sequencing of amplified portion of mt DNA D-loop was done. Sequences were analyzed with the help of software blast2sequence. Single Nucleotide Polymorphisms (SNPs) were identified and comparison of 52 mitochondrial DNA haplotypes of all buffalo breeds was done. Genetic distance and identity between five buffalo breeds were calculated and phylogenetic tree was constructed using BioEdit and MEGA 4.1 softwares showing the relationships between different haplotypes. Domestication events were also observed through network analysis. For further confirmation of the genetic structure of buffalo breeds 8 dye labeled microsatehhite markers (recommended by ISAG) were used and genotyping was done. Results were analyzed with the help of different softwares. Genetic diversity, Allele frequencies, observed and expected homozygosity and heterozygosity, Hardy Weinberg equilibrium, F-Statistics and Gene Flow for all Loci, population's dendogram, Neis genetic identity and genetic distance/ diversity was calculated. This work provided the genetic data which is very helpful for determining the genetic diversity of buffalo population, breed identification, animal forensic and paternity cases and making effective breeding policies and conservational activities in future.

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