Application of Polymerase Chain Reaction Restriction Fragment Length Polymorphism Technique in Determining the Identity of Several Marine Species in Seafood Products

This study describes an investigation on the application of Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) nucleic acid based technique, as a routine analytical technique to generate DNA fingerprints for 24 fresh marine samples. Data obtained from the DNA fingerprin...

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Main Author:	Lim, Sor Sing
Format:	Thesis
Language:	English English
Published:	2005
Subjects:	Polymerase chain reaction - Marine animals
Online Access:	http://psasir.upm.edu.my/id/eprint/178/1/549023_FBSB_2005_33.pdf
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id	my-upm-ir.178
record_format	uketd_dc
spelling	my-upm-ir.1782013-05-27T06:46:16Z Application of Polymerase Chain Reaction Restriction Fragment Length Polymorphism Technique in Determining the Identity of Several Marine Species in Seafood Products 2005-10 Lim, Sor Sing This study describes an investigation on the application of Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) nucleic acid based technique, as a routine analytical technique to generate DNA fingerprints for 24 fresh marine samples. Data obtained from the DNA fingerprints were used to identify 12 processed marine samples. Samples representative of various species and marine products submitted to different processing conditions were selected to verify the applicability of the techniques. A specific part of mitochondrion (mt) genome, cytochrome b (cytb) gene with 359 base pair (bp) fragment was successfully amplified from all the investigated samples by using ‘universal’ primer pairs. The obtained fragment of cytb gene (359 bp) was digested with different Restriction Endonuclease (RE) resulting in sample specific Restriction Fragment Length Polymorphism (RFLP). A total of 14 fishes, 7 prawns and 3 crabs with the exception of Doublelined tonguesole (Paraplagusia bilineata), Rainbow sardine (Dussumieria acuta), Western king prawn (Metapenaeus latisulcatus), Sharp-rostrum prawn (Parapenaeopsis hardwickii), Giant freshwater prawn (Macrobrachium rosenbergii), Affluent prawn (Thenus orientalis), Giant tiger prawn (Penaeus semisulcatus), Indo-pacific swamp crab (Scylla serrata), Red and Blue swimming crab (Solenocera subnuda) could be differentiated using RE HaeIII, MboII, FokI, and MspI followed by agarose gel electrophoresis. For processed marine samples, only four out of twelve were successfully identified. The DNA of unidentifiable samples may have been degraded during the steps of processing. The RFLP patterns obtained are conclusive even in the mixture of Western king prawn (Metapenaeus latisulcatus) and African threadfin (Alexis alexandrinus) at a ratio of 1:100. Results of this study suggest that the PCR-RFLP based on cytb gene shows a reproducible, rapid and simple method for simultaneous identification of marine samples. Polymerase chain reaction - Marine animals 2005-10 Thesis http://psasir.upm.edu.my/id/eprint/178/ http://psasir.upm.edu.my/id/eprint/178/1/549023_FBSB_2005_33.pdf application/pdf en public masters Universiti Putra Malaysia Polymerase chain reaction - Marine animals Faculty of Biotechnology and Biomolecular Sciences English
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
topic	Polymerase chain reaction - Marine animals
spellingShingle	Polymerase chain reaction - Marine animals Lim, Sor Sing Application of Polymerase Chain Reaction Restriction Fragment Length Polymorphism Technique in Determining the Identity of Several Marine Species in Seafood Products
description	This study describes an investigation on the application of Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) nucleic acid based technique, as a routine analytical technique to generate DNA fingerprints for 24 fresh marine samples. Data obtained from the DNA fingerprints were used to identify 12 processed marine samples. Samples representative of various species and marine products submitted to different processing conditions were selected to verify the applicability of the techniques. A specific part of mitochondrion (mt) genome, cytochrome b (cytb) gene with 359 base pair (bp) fragment was successfully amplified from all the investigated samples by using ‘universal’ primer pairs. The obtained fragment of cytb gene (359 bp) was digested with different Restriction Endonuclease (RE) resulting in sample specific Restriction Fragment Length Polymorphism (RFLP). A total of 14 fishes, 7 prawns and 3 crabs with the exception of Doublelined tonguesole (Paraplagusia bilineata), Rainbow sardine (Dussumieria acuta), Western king prawn (Metapenaeus latisulcatus), Sharp-rostrum prawn (Parapenaeopsis hardwickii), Giant freshwater prawn (Macrobrachium rosenbergii), Affluent prawn (Thenus orientalis), Giant tiger prawn (Penaeus semisulcatus), Indo-pacific swamp crab (Scylla serrata), Red and Blue swimming crab (Solenocera subnuda) could be differentiated using RE HaeIII, MboII, FokI, and MspI followed by agarose gel electrophoresis. For processed marine samples, only four out of twelve were successfully identified. The DNA of unidentifiable samples may have been degraded during the steps of processing. The RFLP patterns obtained are conclusive even in the mixture of Western king prawn (Metapenaeus latisulcatus) and African threadfin (Alexis alexandrinus) at a ratio of 1:100. Results of this study suggest that the PCR-RFLP based on cytb gene shows a reproducible, rapid and simple method for simultaneous identification of marine samples.
format	Thesis
qualification_level	Master's degree
author	Lim, Sor Sing
author_facet	Lim, Sor Sing
author_sort	Lim, Sor Sing
title	Application of Polymerase Chain Reaction Restriction Fragment Length Polymorphism Technique in Determining the Identity of Several Marine Species in Seafood Products
title_short	Application of Polymerase Chain Reaction Restriction Fragment Length Polymorphism Technique in Determining the Identity of Several Marine Species in Seafood Products
title_full	Application of Polymerase Chain Reaction Restriction Fragment Length Polymorphism Technique in Determining the Identity of Several Marine Species in Seafood Products
title_fullStr	Application of Polymerase Chain Reaction Restriction Fragment Length Polymorphism Technique in Determining the Identity of Several Marine Species in Seafood Products
title_full_unstemmed	Application of Polymerase Chain Reaction Restriction Fragment Length Polymorphism Technique in Determining the Identity of Several Marine Species in Seafood Products
title_sort	application of polymerase chain reaction restriction fragment length polymorphism technique in determining the identity of several marine species in seafood products
granting_institution	Universiti Putra Malaysia
granting_department	Faculty of Biotechnology and Biomolecular Sciences
publishDate	2005
url	http://psasir.upm.edu.my/id/eprint/178/1/549023_FBSB_2005_33.pdf
_version_	1747810167341187072

Application of Polymerase Chain Reaction Restriction Fragment Length Polymorphism Technique in Determining the Identity of Several Marine Species in Seafood Products

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