scholarly journals Identification and differentiation among chicken’s, duck’s, quail’s, rabbit’s and turkey's meat using PCR-RFLP technique

2015 ◽  
Vol 31 (1) ◽  
pp. 101-108 ◽  
Author(s):  
S.M. Abdel-Rahman ◽  
A.M. Elmaghraby ◽  
A.S. Haggag
Keyword(s):  
Mitochondrial Dna ◽  
Cytochrome B ◽  
Fragment Length ◽  
Restriction Analysis ◽  
Restriction Enzymes ◽  
Pcr Product ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Species Specific ◽  
Rflp Assay

PCR-RFLP technique was developed for identification and differentiation among chicken?s, duck?s, quail?s, rabbit?s and turkey's meat. DNA from small amount of muscles (0.05 g) was extracted and a region of mitochondrial DNA (cytochrome-b gene) in chicken, duck, quail, rabbit and turkey was amplified by PCR. Fragment length of the PCR product was 371 bp in chicken, 374 bp in duck and rabbit and 377 bp in both quail and turkey. Six nucleotides different makes it difficult to differentiate among these five species-specific meat. For differentiation, three different restriction enzymes (DdeI, MspI and TaqI) were used to digest the PCR products. Restriction analysis showed difference among chicken?s, duck?s, quail?s, rabbit?s and turkey's meat. Where, DdeI yielded two fragments (291 and 83 bp) only in rabbit?s meat. MspI yielded three fragments (221, 85 and 65 bp) in chicken?s meat and two fragments (290 and 87 bp) in both quail?s and turkey's meat. TaqI yielded three fragments (146, 134 and 94 bp) in duck?s meat and two fragments (226 and 151 bp) in quail?s meat. The use of Cytb- PCR-RFLP assay allowed a direct and fast authentication and differentiation among chicken?s, duck?s, quail?s, rabbit?s and turkey's meat.

scholarly journals Evidences reveal that cattle and buffalo evolutionary derived from the same ancestor based on cytogenetic and molecular markers

2006 ◽  
Vol 22 (3-4) ◽  
pp. 1-9 ◽  
Author(s):  
S.M. Abdel-Rahman
Keyword(s):  
Cytochrome B ◽  
Fragment Length ◽  
Restriction Analysis ◽  
Pcr Amplification ◽  
Gene Encoding ◽  
Ssr Analysis ◽  
Mitochondrial Cytochrome B ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Species Specific

Muscle-DNA from cattle and buffalo was extracted to amplify the mitochondrial DNA segment (cytochrome b gene) and the gene encoding species-specific repeat (SSR) region. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and SSR techniques were used to identify of species origin. Restriction analysis of PCR-RFLP of the mitochondrial cytochrome b segment and SSR analysis showed no differences between cattle and buffalo. Where, the fragment length (bp) generated by AluI PCR-RFLP were 190, 169 and PCR amplification size of the gene encoding SSR region was 603 bp in both cattle and buffalo. Consequently, finding from this study could be revealed that cattle and buffalo are evolutionary derived from the same ancestor.

PCR-based genetic identification of marlin and other billfish

1998 ◽  
Vol 49 (5) ◽  
pp. 383 ◽  
Author(s):  
B. H. Innes ◽  
P. M. Grewe ◽  
R. D. Ward
Keyword(s):  
Mitochondrial Dna ◽  
Genetic Test ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Genetic Identification ◽  
Dna Molecule ◽  
Pcr Rflp ◽  
D Loop ◽  
Specific Variation ◽  
Species Specific

A genetic test was developed for the identification of the six species of billfish found in Australian waters (black marlin, Indo–Pacific blue marlin, striped marlin, Indo–Pacific sailfish, shortbill spearfish and broadbill swordfish). The test was based on the PCR–RFLP analysis of a 1400 bp region of the mitochondrial DNA molecule, the d-loop, using four restriction enzymes (Hinf I, Rsa I and Sau3A I andTaq I). A total of 33 composite haplotypes were observed among 160 fish; all were species-specific. Three of the species—black marlin, striped marlin and broadbill swordfish—showed sufficient intra-specific variation to be useful in population structure analyses.

scholarly journals The use of cytochrome b and ryanodine polymorphism to identify DNA of animal and human origin

2019 ◽  
Author(s):  
Małgorzata Natonek-Wiśniewska ◽  
Anna Radko
Keyword(s):  
Cytochrome B ◽  
Red Deer ◽  
Dna Analysis ◽  
Roe Deer ◽  
Restriction Analysis ◽  
Dna Profile ◽  
Applied Method ◽  
Gene Coding ◽  
Pcr Product ◽  
Pcr Products

The aim of this study was to determine a match between DNA recovered from evidence material, such as knocked down red deer, and from comparative material in form of two brown traces on the bonnet of a car driven by a person suspected of knocking down the animal. The spots coming from the car provided no DNA profile, which questioned that they originated from a red deer and ruled out performance of a comparative DNA analysis. For this reason, the material obtained from the blood smear was analyzed for species identification. The method applied can discriminate between cattle, red deer and roe deer based on restriction analysis (Tsp509I) of PCR product (195bp), obtained by amplifying a fragment of the cytochrome b coding gene. Because the obtained restriction profile confirmed the match with red deer DNA for one trace, and in the second case ruled out that the biological traces originated from the species mentioned above, the PCR products were subjected to sequencing. In both cases, 195bp PCR products that were 98% homologous with red deer DNA sequence-NC_007704.2-trace1 and with the gene coding for the human ryanodine receptor-NC_008799.2-trace2. The quantity and quality of DNA obtained from the traces collected from the car bonnet did not allow confirmation of the involvement of a specific animal in the event, but the applied method made it possible to determine the species from which the obtained traces originated. Furthermore, the applied method, which was used earlier to determine cervine DNA, was successfully used to detect human DNA.

scholarly journals Polymorphism, Allelic and Genotypic Frequencies of κ-Casein and β-LG genes in Egyptian Buffaloes

2020 ◽  
Vol 12 (4) ◽  
pp. 283
Author(s):  
Z. M. Al-Shawa ◽  
M. F. El-Zarei ◽  
A. A. Ghazy ◽  
M. A. Ayoub ◽  
S. M. Merdan ◽  
...  
Keyword(s):  
Genetic Parameters ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Pcr Product ◽  
Heterozygosity Excess ◽  
Pcr Rflp ◽  
Native Populations ◽  
Egyptian Buffalo ◽  
Rflp Assay ◽  
Geographical Locations

With a view to detecting the genotypes of both κ-CN and β-LG genes in native populations of Egyptian buffalo using PCR-RFLP technique, 80 randomly, individuals were selected from five geographical locations of some Egyptian provinces. Also, to estimate the population genetic parameters such as, allelic and genotypic frequencies, heterozygosity, and inbreeding coefficient (FIS) of these studied genes. For genotyping, 453 bp PCR product of κ-CN was digested with AcuI and HpyCH4IV (Isoschizomer for MaeII) restriction enzymes while the 247 bp PCR product of β-LG was digested with HaeIII restriction enzyme. PCR-RFLP results discovered polymorphism at the level of κ-CN gene in all studied Egyptian buffaloes with two distinct alleles “A” and “B”. PCR-RFLP analysis for κ-CN gene using both restriction enzymes successfully detected that polymorphic status of the studied populations. We recommended using AcuI enzyme which was more capable for differentiating between homozygous (17%) and heterozygous (83%) individuals than HpyCH4IV enzyme which defined only 4% of homozygous individuals and the remaining was heterozygous (96%) individuals. Existence of heterozygosity excess in all studied populations referred to higher degree of genetic variability between individuals within these populations. On contrary, results of PCR-RFLP at the level of β-LG gene revealed a monomorphic pattern of Egyptian buffaloes and genotyped as “AA” animals which signified that PCR-RFLP assay with HaeIII enzyme for β-LG gene failed to discover any evidence of polymorphism in Egyptian buffalo under the circumstances of this study or all studied animals possess only one allele.

Mitochondrial DNA and isozyme electrophoretic analyses of the endangered Acadian whitefish, Coregonus huntsmani Scott, 1987

1991 ◽  
Vol 69 (2) ◽  
pp. 311-316 ◽  
Author(s):  
L. Bernatchez ◽  
T. A. Edge ◽  
J. J. Dodson ◽  
S. U. Qadri
Keyword(s):  
Mitochondrial Dna ◽  
Genetic Model ◽  
Restriction Analysis ◽  
Restriction Enzymes ◽  
Electrophoretic Analysis ◽  
The Other ◽  
Lake Whitefish ◽  
Mitochondrial Genotype ◽  
Species Specific ◽  
Mtdna Restriction Analysis

Electrophoretic analysis of isozymes and mitochondrial DNA (mtDNA) restriction analysis were used to study the genetic divergence between the Acadian whitefish, Coregonus huntsmani, and members of the subgenera Coregonus (lake whitefish, C. clupeaformis) and Leucichthys (Arctic cisco, C. autumnalis, and lake cisco, C. artedii). Results obtained from both studies demonstrated that the Acadian whitefish is genetically highly distinct from the other coregonines examined. mtDNA restriction analysis revealed that the Acadian whitefish possesses a unique mitochondrial genotype which is divergent from that of the two cisco species or lake whitefish. Twelve of 13 restriction enzymes used were informative in distinguishing the Acadian whitefish from the other species, and species-specific fragment patterns were observed for 10 enzymes. In isozyme analysis of five loci, the Acadian whitefish was monomorphic at two loci for alleles not found in lake whitefish. Acadian whitefish also possessed an additional isozyme at another locus that was not found in lake whitefish and Arctic cisco specimens. This isozyme is unknown from the genetic model for lake whitefish at this locus. These results provided useful genetic markers to identify the Acadian whitefish. They emphasize that the extinction of the species would represent a major loss of both genetic diversity and potential information concerning the contentious phylogeny of coregonine fishes.

scholarly journals Identification of restriction enzyme in the FSHR gene of indonesian local cattle

2021 ◽  
Vol 888 (1) ◽  
pp. 012024
Author(s):  
P W Prihandini ◽  
A Primasari ◽  
M Luthfi ◽  
D Pamungkas ◽  
A P Z N L Sari ◽  
...  
Keyword(s):  
Restriction Enzyme ◽  
Restriction Site ◽  
Follicle Stimulating Hormone ◽  
Restriction Enzymes ◽  
Future Studies ◽  
Fshr Gene ◽  
Pcr Rflp ◽  
Mapping Analysis ◽  
Pcr Products ◽  
Enzyme Mapping

Abstract The restriction enzyme is important for genotyping using the PCR-RFLP technique. Therefore, this study aims to identify the restriction enzyme mapping in the partial sequence of the follicle-stimulating hormone receptor (FSHR) gene in Indonesian local cattle. A total of 29 samples sized 306 bp, were aligned with Genbank sequence acc no. NC_032660, resulting three polymorphic sites, namely g.193G>C, g.227T>C, and g.275A>C. Furthermore, the restriction mapping analysis using the NEBcutter program V2.0 showed that no enzyme recognized the SNP g.275A>C, while the SNP g.193G>C and g.227T>C were identified by the AluI and MscI enzymes, respectively. The AluI enzyme cuts at two positions (193 bp and 243 bp) in the G allele sample producing three fragments namely 50 bp, 63 bp, and 193 bp, meanwhile, in the C allele, the AluI cuts only in position 243 bp, hence, the fragment products are 63 bp and 243 bp. In contrast, the MscI enzyme was only recognized in the T allele, producing fragments sized 77 bp and 229 bp but failed to identify the restriction site along with the PCR products in the C allele. Based on the results, the SNPs (g.193G>C and g.227T>C) and restriction enzymes (AluI and MscI) are applicable for genotyping local Indonesian cattle using the PCR-RFLP technique in future studies.

Comparative Fingerprinting Analysis ofCampylobacter jejuni subsp. jejuni Strains by Amplified-Fragment Length Polymorphism Genotyping

2000 ◽  
Vol 38 (9) ◽  
pp. 3379-3387 ◽  
Author(s):  
Bjørn-Arne Lindstedt ◽  
Even Heir ◽  
Traute Vardund ◽  
Kjetil K. Melby ◽  
Georg Kapperud
Keyword(s):  
Length Polymorphism ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Amplified Fragment Length Polymorphism ◽  
Epidemiological Surveillance ◽  
Aflp Analysis ◽  
Polymorphism Analysis ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Ofcampylobacter Jejuni

Amplified-fragment length polymorphism (AFLP) analysis with the endonucleases BglII and MfeI was used to genotype 91 Campylobacter jejuni subsp. jejunistrains from outbreaks and sporadic cases. AFLP-generated fragments were labeled with fluorescent dye and separated by capillary electrophoresis. The software packages GeneScan and GelCompar II were used to calculate AFLP pattern similarities and to investigate phylogenetic relationships among the genotyped strains. The AFLP method was compared with two additional DNA-based typing methods, pulsed-field gel electrophoresis (PFGE) using SmaI and restriction fragment length polymorphism analysis on PCR products (PCR-RFLP) of theflaA and flaB genes. We found that AFLP analysis of C. jejuni strains is a rapid method that offers better discriminatory power than do both PFGE and PCR-RFLP. AFLP and, to a lesser extent, PCR-RFLP could differentiate strains within the same PFGE profiles, which also makes PCR-RFLP an alternative to PFGE. We were able to clearly distinguish 9 of 10 recognized outbreaks by AFLP and to identify similarities among outbreak and sporadic strains. Therefore, AFLP is suitable for epidemiological surveillance ofC. jejuni and will be an excellent tool for source identification in outbreak situations.

Cloth-Based Hybridization Array System for Expanded Identification of the Animal Species Origin of Derived Materials in Feeds

2007 ◽  
Vol 70 (12) ◽  
pp. 2900-2905 ◽  
Author(s):  
JOHANNA MURPHY ◽  
JENNIFER ARMOUR ◽  
BURTON W. BLAIS
Keyword(s):  
Mitochondrial Dna ◽  
Dna Sequences ◽  
Microscopic Examination ◽  
Animal Species ◽  
Specific Detection ◽  
Animal Feeds ◽  
Species Origin ◽  
Pcr Products ◽  
Species Specific ◽  
Immunoenzymatic Assay

A cloth-based hybridization array system (CHAS) previously developed for the detection of animal species for which prohibited materials have been specified (cattle, sheep, goat, elk, and deer) has been expanded to include the detection of animal species for which there are no prohibitions (pig and horse) in Canadian and American animal feeds. Animal species were identified by amplification of mitochondrial DNA sequences by PCR and subsequent hybridization of the amplicons with an array of species-specific oligonucleotide capture probes immobilized on a polyester cloth support, followed by an immunoenzymatic assay of the bound PCR products. The CHAS permitted sensitive and specific detection of meat meals from different animal species blended in a grain-based feed and should provide a useful adjunct to microscopic examination for the identification of prohibited materials in animal feeds.

scholarly journals Identification of Pneumococcal Serotypes by PCR–Restriction Fragment Length Polymorphism

Diagnostics ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 196 ◽  
Author(s):  
García-Suárez ◽  
González-Rodríguez ◽  
Cima-Cabal ◽  
Yuste ◽  
Vazquez ◽  
...  
Keyword(s):  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Dna Sequences ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Pcr Rflp ◽  
Restriction Fragment Length

Streptococcus pneumoniae shows more than 90 capsular serotypes that can be distinguished by their reactivity against antisera. The main objective of this work was the development of a molecular method for serotyping without the use of antisera. A computer program containing an algorithm was used to search in a database for potentially useful enzymes for Restriction Fragment Length Polymorphism-RFLP typing, in order to maximize the discrimination between different serotypes. DNA sequences of 90 serotypes for the region between dexB and aliA genes were compiled, and a computer screening of restriction enzymes was performed. The wzg–wzh–wzd–wze region and Sse9I restriction predicted unique PCR-RFLP patterns for 39 serotypes and eight serogroups. A second restriction enzyme resolved fragment specific patterns for 25 serotypes. The method was tested with 98 serotype-unknown clinical isolates. PCR-RFLP analysis deduced correct serotypes that were confirmed by Quellung reaction for 78.5% of the isolates.

Variation in the mitochondrial DNA of the potential biological control agent Chondrostereum purpureum

2000 ◽  
Vol 77 (10) ◽  
pp. 1490-1498 ◽  
Author(s):  
Tod D Ramsfield ◽  
Simon F Shamoun ◽  
Zamir K Punja ◽  
William E Hintz
Keyword(s):  
Biological Control ◽  
British Columbia ◽  
Mitochondrial Dna ◽  
New Zealand ◽  
Cytochrome B ◽  
Biological Control Agent ◽  
Gene Diversity ◽  
Restriction Enzymes ◽  
Sample Population ◽  
Chondrostereum Purpureum

The mitochondrial DNA (mtDNA) of Chondrostereum purpureum (Pers.:Fr.) Pouzar was extracted and purified, and the size ranged from 51.8 to 66.4 kb. One isolate each from British Columbia, Alberta, Finland, the Netherlands, and New Zealand were found to have identical BamHI mtDNA restriction patterns, resulting in a mitochondrial genome of 63.8 kb. An additional isolate from British Columbia and one from Switzerland had different banding patterns, however, resulting in mitochondrial genomes of 66.4 kb and 51.8 kb, respectively. A sequence-characterized amplified region (SCAR) assay, based on a polymerase chain reaction, was developed to rapidly screen a larger population of 84 isolates from North America, Europe, and New Zealand. Two SCARs, one encoding the NADH 4 gene (3 kb) and the second encoding the ATPase VI and cytochrome b genes (5.1 kb), were digested with 24 restriction enzymes. There were no polymorphisms in the NADH 4 containing SCAR, while a single polymorphism was detected by NsiI in the ATPase VI - cytochrome b containing SCAR. Two mitochondrial haplotypes that were distributed throughout the sample population were thus identified. The coancestry coefficient (<$Q7A0D00000010446D80BFFEFF88A524F5343905055B98C420120907B4DDA9ECB1F0>) for all subpopulations of the sample population was calculated to be 0.0353. The level of gene diversity in the mtDNA ofC. purpureum suggested that the chance introduction of novel mitochondrial genes following biological control applications of the fungus is relatively low.

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