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1. Study Of Genetic Association Of Pax Gene To Waardenburg Syndrome In Pakistani Patients

by Huma Tabassam | Dr. Ali Raza Awan | Miss. Sehtish Faryal | Mrs. Shagufta.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2011Dissertation note: Waardenburg syndrome is an auditory-pigmentary disorder that combines clinical features of pigmentary abnormalities of the skin, hair and irides, sensorineural hearing loss, and Hirschsprung disease. Patients with WS have been shown to have mutations in the PAX gene as well as in other genes. In the present study, the locus specific polymorphisms of human PAX gene isolated from healthy and diseased Pakistani individuals was investigated for genetic association of the polymorphism with the Waardenburg syndrome. Mutation in PAX gene was identified from Pakistani patients with Waardenburg syndrome. For the purpose, blood samples of patients suffering from Waardenburg syndrome were collected from different hospitals. DNA was isolated from WBCs suspended in the preserved samples using standard organic DNA extraction protocol. Primers were designed using Primer3 programme. PCR conditions were optimized and mutation discovery was performed on all DNA samples. Analysis of the sequences and mutations was done with the help of appropriate bioinformatics softwares. Analysis of the variable sites revealed T?C transitions, (mutations) at position number 244 was found in exon 2 of the gene. All 17 patients exhibit this mutation at the same position. Results of normal patients where there was no change found and PAX3 gene is not mentioned as it was not significant to mention. PAX gene has not been studied in the Pakistani patients earlier. The mutational investigation and association study will help to understand the genetic basis of the Waardenburg syndrome not only in Pakistan but it will also contribute to the global efforts to understand the human genetics. Availability: Items available for loan: UVAS Library [Call number: 1359,T] (1).

2. Association Of Genetic Polymarphism Of Cyp 2D6 Gene With Generalized Tonic Clonic Seizures In Pakistani Ptients

by Rana Manzoor Ahmad | Dr. Ali Raza Awan | Prof. Dr.Masroor Elahi Babar.

Material type: book Book; Format: print Publisher: 2011Dissertation note: Epilepsy is chronic neurological disorder in which hyperexcitibilty of neurons cause seizures. It is a serious disorder as there is an association between the increased mortality and epilepsy. The etiology of epilepsy can be genetic. There are two main types of epilepsy, partial and generalized. These two types are further categorized into different type of seizures. Its prevalence is more in developing countries like Pakistan. The generalized tonic clonic seizure (GTCS) is a type of epilepsy prevelant in Pakistan. Cytochrome P450 (CYP2D6) enzyme has been reported to be associated with GTCS. CYP2D6 is encoded by a 4.6 kb gene named as CYP2D6. This is a highly polymorphic gene having 09 exons. In this study CYP2D6 gene (exon 1-5) was characterized for polymorphism and the polymorphism was evaluated for asssociation with GTCS in Pakistani patients. Patient data and blood samples of different epilepsy patients were collected. DNA was isolated by inorganic method. PCR amplification was used for amplification of CYP2D6 (exon 1-5) and sequencing was performed on ABI 3130 XL Genetic analyzer. Two mutations, 214 G>C and 232 G>C in intron1 of CYP2D6 gene have been found. These mutations were only found in Pakistani patients suffering with GTCS. Absence of these mutations in 10 healthy individuals (control group) confirmed association of these mutations with GTCS. This outcome of study will help to add information in international gene data. The mutations found in this study will also lead to gene therapy of GTCS, genetic counseling and develop prenatal diagonastic tests. Availability: Items available for loan: UVAS Library [Call number: 1390,T] (1).

3. Association Of Katg Gene With Isoniazid Resistance In Multiple Drug Resistant Tuberculosis

by Farouk Qamar Malik | Dr. Ali Raza Awan | Prof. Dr.Masroor Elahi Babar.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2012Dissertation note: TB has been announced as a global emerg~ncy of this millennium. It is one of the leading causes of death among adults due to a single infectious agent. Pakistan is sixth among the twenty two Eastern Mediterranean Region countries with the highest burden of disease which is approximately 181 per 100,000. The emergence of drug resistant MTB poses a serious threat to the ongoing efforts to control the disease epidemic. Drug resistance to the first line drugs such as INH and RIF needs to be investigated. In this respect the role of various genes conferring resistance should be studied to find better treatment alternatives. The current practice of drug sensitivity testing requiring approximately three weeks (total turnaround time for MTB culture and sensitivity is around 90 days) is time consuming and a major cause of treatment delay. In this research sputum samples were collected in wide mouth transparent containers from suspected TB patients. After decontamination samples were inoculated onto LJ medium. The colonies grown on the slopes were identified as MTB by standard biochemical test. Isolates were tested on LJ medium for in vitro DST (Drug Sensitivity Testing). MDR was described as resistance to INH and RIF with or without resistance to other drugs. DNA was extracted from the grown samples using kit method. After extraction of DNA, the region from base 2714 to 3232 of katG gene was amplified through peR and the amplified products were sequenced. Analysis of the DNA sequences and mutations was done with the help of BLAST - alignment software. A total of 24 MDR MTB samples were sequenced. Sequence analysis revealed the reported mutation Ser - Thr in katG codon 315 in five samples (21 percent of the total sample size). In this study, an authentic molecular analysis (test) was developed and validated for identification of INH resistant strains in Pakistani population. By studying genetic mutations in katG gene and its association With INH resistance, an alternative can be provided whereby specimens can be tested for these mutations and timely decisions taken. This will not only save the patients from unnecessary treatment delays but will also prevent the administration of drugs to which MTB is resistant and in the long run decrease drug resistance and disease burden. Availability: Items available for loan: UVAS Library [Call number: 1402,T] (1).

4. Dna Barcoding Of Pakistan Avian Families (Sturnidae & Columbidae)

by Emma Umar | Dr. Ali Raza Awan | Dr. Muhammad | Ms. Saeeda Kalsoom.

Material type: book Book; Format: print Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1515,T] (1).

5. Genetic Characterization Of Livestock Species Of Pakistan Through Dna Barcoding

by Madiha Booter | Dr.Ali Raza Awan | Dr. Abu saeed | Dr. Muhammad Imran.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2013Dissertation note: The interaction of livestock with ecosystem plays a vital role in sustainability of life. The demand of livestock products is rising day by day which is changing the relationship between livestock and natural resources. Livestock animals are playing a major role towards domestication and also contributing to fulfill human needs through meat and milk production for food industry, which generate big revenues. Pakistan is blessed with the world's best livestock species and there is a need to establish a well characterized system for the classification and identification of these important livestock species. Mitochondrial DNA is of small size, constitutes a small fraction of the total of cell's genome and due to high rate of mutation, it is considered to be an ideal model to study evolutionary relationships. DNA barcoding is being used to characterize animals by using a standard region of mitochondrial DNA as a molecular marker. The study is designed to develop the DNA barcode for genetic characterization of livestock species of Pakistan which includes sheep, goat, cow, buffalo and camel. Blood samples were collected from the selected livestock species. Primers were designed using primer designing free-ware software. The amplified PCR products weresequenced in both orientations by chain termination method. For data analysis,Chromas was used to read sequencing results. To study variation in all sequenced data, alignment tools were used from NCBI. Theblastnalignment tool available at NCBI is more reliable to give authentic results.The alignment results showed 100% homology with the reference sequences (No SNP or mutation was identified). The results can further be validated with the help of mass level sampling to rationalize the study at population level.Phylogenetic analysis indicated that COIDNA barcode region can be used to discriminate unknown samples of any of the species under consideration. The COIgene successfully cladded already reported sequences of the same species. This study provided genetic data which help in species identification, to assess evolutionary pattern and genetic diversity. So, it will also be helpful to monitor legal or illegal trade of livestock species and to identify processed and unprocessed meat for quality assurance. Establishment of an elaborated DNA barcode system for livestock species will help to start taxonomic investigation and will lead towards to identify many new mammalian species of Pakistan. Availability: Items available for loan: UVAS Library [Call number: 1752,T] (1).

6. Genetic And Evolutionary Characterization Of Pakistani Pigeons And Parrots Through Mitochondrial D-

by Sehrish firyal | Dr. Ali raza awan | Prof, Dr. Aftab | Prof, Dr. Tahir yaqub.

Material type: book Book; Format: print Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1873,T] (1).

7. Phylogenetic Analysis Of Major Fresh Water Carps Of Pakistan Through DNA Barcoding

by Madeeha Awan (2012-VA-650) | Dr. Ali Raza Awan | Dr.Sehrish Firyal | Dr. Muhammad Tayyab.

Material type: book Book; Literary form: not fiction Publisher: 2014Dissertation note: Pakistan is bestowed with the land of geological and topographic diversity. The ecological variation is uniformly reflected in all water lands of the country. Pakistan has significantly huge natural inland water resources in the form of ocean, rivers, networks of canals and lakes (Mirza and Rafique 1994). The country is blessed with one of the largest freshwater resources in the world correspondingly large number of freshwater living vertebrates is available from which fishes are quite significant considering the ecological balance and its consumption as food. It is one of the food sources which solely provide all the essential nutrients, minerals and high quality protein which is not common from other food items (Muhammad Rafique 2007). Out of 33,100 fish species identified worldwide as per Fish Base organization report published in April 2015 (http://www.fishbase.org). Out of 233 (indigenous and exotic) freshwater fish species, 78 economically important indigenous fish species are available in the water bodies of the Pakistan. According to this report fishes are the largest vertebrate group, constituting about 50% of all vertebrate species. Systematically fishes are widely spread in nature, ranging from prehistoric jawless fishes to cartilaginous fishes and also from old to current day bony fishes. The taxonomic placement of these fishes shows their belonging to class Actinopterygii, sub-class Teleostei, 3 cohorts, 6 superorders, 13 orders, 30 families and 86 genera (Rafique 2007; Rafique and Khan 2012). Demand of fish is increasing day by day not only being the naturally available source of food rather the health benefits associated with its consumption. This necessitates to develop a more efficient and sustainable system to increase their growth. DNA based technologies are being competently employed in aquaculture production fields for pedigree information. Introduction 2 Moreover, tagging each fish individually is not an easy task so these DNA based methods help in avoiding intrusion of environmental factors which may result from raising fish families in separate reservoirs (Martinez 2007). Fish identification has been traditionally based on phenotypic features. However, due to high multiplicity and morphological similarity, in many cases, fish at its different developmental stages are hard to be identified by relying only on morphological characteristics (Victor et al. 2009). For phylogenetic studies of the animals the use of mt-DNA is very common and reliable compared to nuclear DNA due to its high evaluation capabilities, which results in gathering of differences even between closely related species (Moore 1995; Mindell et al. 1997).“Bar-coding gap" is the name given to the property that is inter-specific variation in this region is markedly higher than intra-specific variability (Hebert et al. 2003). Approximately each and every animal contain 13 protein-coding genes (PCGs) as an essential component of their mt-genome (mitochondrial genome), which helps in encoding of several proteins responsible for the oxidative phosphorylation machinery (Richly A et al. 2004, Song H et al. 2008). Being maternally inherited, mt-DNA is better as compared to genomic DNA such as quick evolution, less exposure to recombination, high copy number, high conservation, little duplications and negligible intergenic regions (Waugh J 2007, Xu J 2005). Clonal inheritance is the main property which makes it more worthy and suitable marker in comparison with other available molecular bio-diversity tools (Galtier et al. 2009). DNA barcoding is one of the taxonomic tools. It is being used to distinguish animal species based on the small segment of their genome such as mitochondrial DNA, designated as an identification tag or barcode of particular species (Herbert et al. 2003). Identification of species using DNA barcoding is quite debatable. Still many researchers consider it as a reliable Introduction 3 basic tool to ascertain the genetic characterization of diverse eukaryotic species, especially after establishment of the Consortium for the Barcode of Life (CBOL) in 2004 [http: //www. barcodeoflife.org/]. Ideally DNA barcoding should provide quick, reliable and cost effective species identification, even to those user who has little or negligible knowledge of taxonomy (Herbert et al. 2003, Hajibabaei M et al. 2005, Herbert et al. 2005). Identification of unknown source is possible by using distance based tree which can be created by comparing unidentified sequences against retrieved known sequences of different species (Hebert et al. 2003, 2004a, 2004b). DNA barcoding identification system has been recognized universally as standardized method to recognize species and unveil their genetic diversity (Herbert et al. 2003; Herbert et al. 2004). The ideal DNA barcoding is robust, with conserved, universal primer binding sites, reliable DNA amplification and sequencing. From whole mitochondrial genome, Cytb (Cytochrome b) is considered as one of the most promising gene due to its function and structure, even it is composed of both conserved and rapidly evolving regions which are more related to evolutionary studies (Farias et al. 2001). To identify unknown or ambiguous species it is considered more reliable as it contain sequences which provide the specific information about particular species (Parson W et al. 2000a, 2000b). It is also one of the most useful genetic marker to identify the linkage within families and genera (Meyer 1994; Teletchea 2009). Cytb gene is involved in comparative studies which results in development of new classification schemes and been used to assign a genus to a newly described species as well as improve the understanding of evolutionary relationships of genra (Castresana 2001). Introduction 4 One of the core objectives of this study is to identify and classify four freshwater indigenous fish species of Pakistan, which includes Labeo rohita (Rohu), Labeo calbasu (Calbans), Catla catla (Thalla) and Cirrhinus mrigala (Mori) using Cytb gene. Morphologically, Labeo rohita shows compressed body with convex dorsal profile while mouth bears a pair of barbells and fins are gray and orange in color. Catla catla shows compressed body with broad head. Mouth is wide with thick lower lip. Labeo Calbasu`s dorsal profile is more convex than that of abdomen and two pairs of barbells are present on fringed upper lip. Cirrhinus mrigala has elongated and streamlined body shape which is grayish and silver in color (Bhuiyan AL 1964; Rahman AK 1989). All of these species are found in freshwater bodies mostly lakes, rivers and ponds except Labeo calbasu which is a bottom dweller. These fishes are harvested by using rod and line or by using nets (Talwar PK and Jhingran AG 1991). These fishes are known as major carps and economically very important for the country due to their high consumption as food. These fishes are also used for fish farming due to their greater muscle mass thus also possess very high commercial value for fish farming business. Another objective of this study is to resolve the taxonomic anomalies related to above mentioned species. Selling of fish meat in mislabeled packaging is a serious issue now days. Most commonly Hypophthalmichthys molitrix (silver carp) and Ctenopharyngodon idella (grass carp) are sold under the label of Labeo rohita. DNA barcoding is also helpful in detecting such fraudulent mislabeling. It would be the first study in Pakistan to genetically characterize commercially important fish species. It would help scientists to know about their phylogenetic and taxonomic status and also assist fish fanciers to genuinely identify their species of interest. Identification of fish species is also important for conservation of biodiversity as it helps in preservation and Introduction 5 identification of endangered species by generating their barcodes from even minimal evidence available. This study has paved the way for molecular biologists to study taxonomic ambiguities at sub species level using SNP (Single nucleotide polymorphism) based identifying marker. Availability: Items available for loan: UVAS Library [Call number: 2207-T] (1).

8. Genetic Characterization Of Pakistani Wild Quails Using Mitochondrial Coi Gene

by Wajiha Shakil (2012-VA-817) | Dr. Ali Raza Awan | Dr. Muhammad Yasir Zahoor | Prof. Dr. Tahir Yaqub.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: A latest taxonomic tool termed as DNA barcoding is being used to genetically characterize animals. DNA barcoding is helpful in determining evolutionary relationship among species. Being a small sized genome and agile enough to show rapid mutation, mtDNA has been used as a pertinent marker of molecular biodiversity. DNA barcode works as an efficient tool in food manufacturing industry, diet investigation, forensics analysis, preventing unlawful trade and felonious poaching. The aim of this study was to develop DNA barcode for genetic characterization of Pakistani wild quail. Pakistani wild quail is important due to its demand for eggs, meat production, experimental purposes and gaming as well. Japanese quail was also included in this study because this quail is excessively produced in Pakistani farms. Japanese quail is present throughout the year and is comparatively bigger in size than wild quail. It has longer lifespan; farmers can easily breed this species in farms. It is suitable in poultry due to better meat yield. COI gene (500bp) was used as a molecular marker for identification at species level. DNA was extracted from blood samples of ten wild quails (Coturnix coturnix and fifteen japanese quails (Coturnix japonica). Reported bird universal primers were used to amplify COI region from the extracted mtDNA samples using PCR. Amplicon were then sequenced by Sanger sequencing method (Sanger et al. 1977). Forward and reverse DNA Sequences were aligned with the reference sequence using nucleotide BLAST on NCBI to observe the dissimilarity among the sequences. Consensus sequences generated were used to construct their phylogenetic tree to see their evolutionary relationship with other bird species. Japanese quail which is thought to be domesticated from Japan, its Pakistani population showed close relation with sequences Summary 90 generated in Japan for this particular species. Pakistani wild quail species showed its closest linkage with C. coturnix. In conclusion, COI barcode proved as an authentic tool for species identification and phylogenetic inference of Pakistani wild and farm grown quails. Wild quail species has been characterized using partial COI gene sequences. This study has provided a specific genetic marker which can differentiate Japanese quail from wild quail at molecular level as most of the time both species are confused with each other. It can be helpful to the farmers and bird fanciers because they can select the birds of their choice correctly. This is the first study reporting DNA barcode of this Pakistani quail species. It would help researchers to study about phylogenetic and taxonomic status and also assist quail fanciers and quail farmers to unaffectedly identify their species of interest in farming. Identification of quail species is also important for conservation of biodiversity as it helps in preservation and identification of endangered species by generating their barcodes from even minimal evidence available. Availability: Items available for loan: UVAS Library [Call number: 2311-T] (1).

9. Assessment Of Evolutionary Rate In Different Serotypes Of Foot & Mouth Disease Virus

by Muhammad Farooq (2011-va-823) | Dr. Ali Raza Awan | Prof. Dr. Thair Yaqub | Dr. Muhammad Tayyab.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: FMDV belongs to the family Picornaviridae with seven serotypes around the world. Nevertheless, serotypes prevalent in Asia includes A, O and Asia-1. Because of evolution in genomic sequence of FMDV, it is becoming difficult to control the problem through conventional methods. Changes in the genome can be detected using software through sequence analysis. In the software, evolutionary models are used to measure the evolutionary change for the identification of new sub lineages. In current study genomes sequence data (1998 - 2011) of bovine FMD serotypes (A, O and Asia 1) was collected through NCBI in FASTA format. This data was converted into PHYLIP format. On Dell workstation, with Microsoft Windows 8.1 operating system, BioEdit, TipDate V.1.2 was deployed. Sequence data was aligned through CLUSTAL W algorithm of Multiple Sequence Alignment using BioEdit. Using TipDate, genome sequence data was analyzed using three evolutionary models (F84, HKY and REV) and phylogenetic trees were produced showing evolutionary rate and likelihood ratio of FMDV serotypes O, A and Asia-1.. Results of the current study showed higher values of evolutionary rate in bovine FMD virus which was estimated 7.49 x 10-4 with likelihood value -1429.507680 in serotype A, 2.418 x 10-3 with likelihood -3707.168484 in serotype O and2.16 x 10-3 with likelihood value -3723.344884 in serotype Asia-1, respectively. Markove Reversible Model showed higher rates of evolution in all three serotype with best likelihood values. Phylogenetic results showed higher rate of evolution or substitution in viruses. Furthermore serotypes A, O and Asia-1are mutating with passage of time and new variants are being observed. It was also observed that this evolutionary process is continued in these three serotypes during 1998-2011. This study confirmed the evolutionary changes in FMDV serotypes prevalent in Pakistan during the period 1998 – 2011. This study showed that isolate are evolving with increasing rate. High rate of mutation in Asia-1 was observed than serotype A and serotype O. F84 and HKY85 models produced close results but these models are not identical works on equal and unequal base frequencies. Markove model estimated average base substitution with mutation and depicts good phylogenetic trees of sequence data. Availability: Items available for loan: UVAS Library [Call number: 2372-T] (1).

10. Genetic Identification And Molecular Classification Of Sub-Family Phasianinae Of Pakistani Bird Species Through Dna Barcoding

by Maryem Hussain (2008-VA-349) | Dr. Ali Raza Awan | Dr. Sehrish Firyal | Dr. Muhammad Tayyab.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: DNA barcoding is a precise technique that uses molecular genetics tools for accurate identification, categorizing, relating and separating the phylogenies of species. Being a small sized genome and agile enough to show rapid mutation, mtDNA has been used as a pertinent marker of molecular biodiversity.The aim of this study was to develop DNA barcode for genetic characterization and classification of Sub-family Phasianinaeof Pakistani bird species. Theyhave not been genetically identified yet in Pakistan. It includes birds like domestic chicken(Gallus gallusdomesticus), aseel chicken(Gallus gallusdomesticus strain),blue peafowl(Pavo cristatus), green peafowl (Pavo muticus), white peafowl (Pavo cristatus leuticus), Kalij pheasant (Lophura leucomelanos),monal pheasant (Lophophorus impejanus),koklass pheasant(Purcrasia macrolopha), ring necked pheasant (Phasianus colchicus), Tragopan (Tragopan melanocepals) andred junglefowl (Gallus gallus). These birds are considered an important part of an ecosystembecause they play a significant role in seed dissemination, pollination of plants and disease spread which are the basic constituents of an ecosystem. They are used for food, hunting and entertainment purposes. Mitochondrial geneCytochrome c oxidase subunit 1 (CO1)of 500bps was used as a marker for identification at specie level.Genomic DNA was extracted by each blood and tissue sample of eleven bird species (33 samples). Amplification of CO1 gene was a done by using a universal set of primers (BIRDF1 and BIRDR1)containing region of almost 750 bps (Hebert et al. 2003).Amplicons were purified and sequenced Sanger sequencing method (Sanger et al. 1977). Forward and reverse sequences were analyzed using softwaresEMBOSS merger,ClustalW, BioEdit and nBLAST. Phylogenetic analysis of selected bird species was done. Each sequence was aligned with its reference sequences of CO1 gene present on NCBI. Every nucleotide position which did not align with the reference sequence was studied to identify SNPs. Fixation index (FST) were used to measure species diversity within a same sub population relative to that found in the entire population. Consensus sequences (500bps) generated was used to construct their phylogenetic tree to see their evolutionary relationship with other bird species. All species showed their closest linkage with their respective species. Pakistani population of peafowl and chicken species showed the close relation with same sequences generated in China. Tranopans showed its closest linkage with T. temminckii. In conclusion, seven species ofPhasianinaesub-family of Pakistani bird species was genetically characterized first time in Pakistan by using CO1 as a barcode. It proves that DNA barcoding is an efficient and accurate molecular tool for species identifica¬tion and phylogenetic implication. This study leads to establish a DNA Data Bank that helped scientists to investigate the biodiversity, taxonomic classification, specie identification, in forensic purposes and to study the genetic and phenotypic evolution of these species. DNA barcoding through CO1 gene works as a functional tool for detectingmeat mislabeling and preventing illegitimate trade. This study has established foundations for molecular biologists to study taxonomic uncertainties at sub species level using SNP based identifying marker. It helps in preservation and identification of endangered species by generating their barcodes from even minimal evidence available. Availability: Items available for loan: UVAS Library [Call number: 2376-T] (1).

11. Genetic Identification And Characterization Of Pakistani Birds Of Perdicinae Subfamily (Partridge) Through Dna Barcoding Method

by Asim Iqbal Jutt (2013-VA-557) | Dr. Ali Raza Awan | Dr. Muhammad Wasim | Prof. Dr. Kamran Ashraf.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Pakistani birds of Perdicinae sub family are cage and game birds. Birds includes Altectoris chukar, Ammoperdix heyi, Ammoperdix griseogularis, Francolinus francolinus and Francolins pondicerianus. Traditional methods of identification were based on the phenotypical characterization of birds, which may lead to incorrect identification, so there was need to explore their characters at DNA level for accurate identification and to establish a DNA reference. Birds of sub-family Perdicinae have not been genetically characterized in Pakistan. A new precise method “DNA barcoding” was applied using COI gene of mDNA for authentic identification and classification of these birds. Blood and tissue samples of five species (fifteen samples) were obtained. DNA of each sample was extracted by organic method. Amplification of CO1 gene was done by using a universal set of primers BIRDF1, BIRDR1. Sequence of 450bp were analyzed using bioinformatics softwares. Each sample was aligned with its reference sequence of COI gene available on NCBI. Every nucleotide position which did not align with the reference sequence was studied to identify SNPs. A common phylogenetic tree of all partridges showed that they have common ancestor about 0.7 million year ago, F.francolinus, F.pondicerianus and A.heyi shared a common clade whereas A.chukar made a separate clade from the ancestor. A.heyi and F.pondicerianus showed closed resemblance. It has been proved that DNA barcoding is an efficient and accurate molecular tool for species identification and phylogenetic implication. This study established a DNA Data Bank that helped scientists to investigate the biodiversity, taxonomic classification, species identification and also established foundations for molecular biologists to study taxonomic uncertainties at sub species level using SNP based identifying marker. Availability: Items available for loan: UVAS Library [Call number: 2714-T] (1).



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