Development Of Novel Mtdna Metabarcodes For The Species Differentiation Of Class Aves (Record no. 9774)

000 -LEADER
fixed length control field 03121nam a22002057a 4500
005 - DATE AND TIME OF LATEST TRANSACTION
control field 20161122101858.0
008 - FIXED-LENGTH DATA ELEMENTS--GENERAL INFORMATION
fixed length control field 161122b2016 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 2617-T
100 ## - MAIN ENTRY--AUTHOR NAME
Personal name Syeda Rida Mehak Sherazi (2010-VA-477)
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 Aves
260 ## - PUBLICATION, DISTRIBUTION, ETC. (IMPRINT)
Year of publication 2016.
300 ## - PHYSICAL DESCRIPTION
Number of Pages 87p.;
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 Aves. 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 Aves for different forensic and molecular biodiversity analyses.
Blood/feather/tissue samples were collected from Class Aves (one specimen from every order reported to be present in Pakistan). DNA was extracted from the collected specimens through standard organic method, qualified and quantified and then PCR-amplified using novel universal primers selected from aligned mtDNA sequences originating from all Aves 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 presented as a novel metabarcode (16SrRNA) for species level identification of large number of Avian species
In summary, we present universal method for species classification of Aves using a targeted parallel sequencing approach. Both sequencing and phylogenetic studies experiments confirm
Summary
82
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
700 ## - ADDED ENTRY--PERSONAL NAME
Personal name Dr. M. Yasir Zahoor
700 ## - ADDED ENTRY--PERSONAL NAME
Personal name Mr. Shahid Abbas
942 ## - ADDED ENTRY ELEMENTS (KOHA)
Koha item type Thesis
Holdings
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 2016-11-22 2617-T 2617-T Thesis


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