Development Of Novel Mtdna Metabarcodes For The Species Differentiation Of Class Amphibia (Record no. 13612)
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| fixed length control field | 03179nam a22002057a 4500 |
| 005 - DATE AND TIME OF LATEST TRANSACTION | |
| control field | 20170921111621.0 |
| 008 - FIXED-LENGTH DATA ELEMENTS--GENERAL INFORMATION | |
| fixed length control field | 170921b2017 xxu||||| |||| 00| 0 eng d |
| 041 ## - LANGUAGE CODE | |
| Language code of text/sound track or separate title | eng |
| 082 ## - DEWEY DECIMAL CLASSIFICATION NUMBER | |
| Classification number | 2874-T |
| 100 ## - MAIN ENTRY--AUTHOR NAME | |
| Personal name | Rehmatullah (2011-VA-365) |
| 110 ## - MAIN ENTRY--CORPORATE NAME | |
| Location of meeting | Dr. Muhammad Imran |
| 245 ## - TITLE STATEMENT | |
| Title | Development Of Novel Mtdna Metabarcodes For The Species Differentiation Of Class Amphibia |
| 260 ## - PUBLICATION, DISTRIBUTION, ETC. (IMPRINT) | |
| Year of publication | 2017. |
| 300 ## - PHYSICAL DESCRIPTION | |
| Number of Pages | 72p.; |
| 502 ## - DISSERTATION NOTE | |
| Dissertation note | The Folmer COI mtDNA universal primers that are considered standard for DNA barcoding of life contain so many mismatches against the target sequences of vertebrate origin that they often end in failure to amplify many of vertebrate DNA extractions. This discrepancy favors for the selection and designing of new metabarcode primers that can be used to identify all individuals of vertebrates or at least all individuals represented in a class of Vertebrata such as Class Amphibia. The current study embarks on such an endeavor. In this study development of new mtDNA metabarcode (16SrRNA) that can be used as universal primers to amplify almost all species of Class Amphibia for different forensic and molecular biodiversity analyses. Tissue samples were collected from order Urodela of Class Amphibia (Toads , Bull frog and skittering frogs sample were collected from Punjab, Pakistan). DNA was extracted from the collected specimens through standard organic method, qualified and quantified and then PCRamplified using novel universal primers selected from aligned mtDNA sequences originating from order Urodela mitochondrial DNA genomes submitted to different online sequence databases such as NCBI nucleotide database. The sensitivity of PCR also be assessed using a range of DNA concentrations. The amplified products were sequenced on ABI Genetic Analyzer following Sanger’s dideoxy method of sequencing. The correctness of obtained mtDNA sequences were examined visually in Chromas Lite 2.1 software and then alignment of these sequences were performed against highly similar DNA sequences in NCBI nucleotide databases using BLAST in order to identify origin of unknown mtDNA sequences. With the help of sequencing and phylogenetic studies specificity of the universal primer set confirmed and Summary 67 presented as a novel metabarcode (16SrRNA) for species level identification of large number of Amphibian species. In summary, we present universal method for species classification of Amphibia using a targeted parallel sequencing approach. Both sequencing and phylogenetic studies experiments confirm specificity of universal primer set. Although promising results were obtained with current settings, rapid improvement of bench top instruments will further develop method with less hands-on, fewer sequencing errors and lower detection limit. So, in future, this barcode can be used for species identification in various fields of study such as meat adulteration, illegal trade, food mislabeling and molecular estimation of biodiversity. |
| 650 ## - SUBJECT ADDED ENTRY--TOPICAL TERM | |
| Topical Term | Molecular Biology And Biotechnology |
| 710 ## - ADDED ENTRY--CORPORATE NAME | |
| Corporate name or jurisdiction name as entry element | Dr. M. Yasir Zahoor |
| 710 ## - ADDED ENTRY--CORPORATE NAME | |
| Corporate name or jurisdiction name as entry element | Dr. Amjad Riaz |
| 942 ## - ADDED ENTRY ELEMENTS (KOHA) | |
| Koha item type | Thesis |
| Damaged status | Collection code | Permanent Location | Current Location | Shelving location | Date acquired | Full call number | Accession Number | Koha item type |
|---|---|---|---|---|---|---|---|---|
| Veterinary Science | UVAS Library | UVAS Library | Thesis Section | 2017-09-21 | 2874-T | 2874-T | Thesis |