scholarly journals PCR amplification of species-specific repeat for meat DNA identification via genetic markers in cattle & sheep

2005 ◽  
Vol 21 (1-2) ◽  
pp. 1-12 ◽  
Author(s):  
M.M. Ahmed
Keyword(s):  
Dna Sequences ◽  
Pcr Amplification ◽  
Gene Encoding ◽  
Ssr Analysis ◽  
Sheep Meat ◽  
As Species ◽  
Heat Treated ◽  
Cattle Meat ◽  
Species Specific ◽  
Cattle And Sheep

The designed and evaluated four assays based upon PCR amplification of species-specific repeat (SSR) for detection, identification and authentication of cattle and sheep on the DNA level. SSR primers were applied in the polymerase chain reaction (PCR), the products has been used for the specific identification of cattle and sheep meat. PCR amplification size of the gene encoding SSR region in cattle and sheep meat was 603 bp and 374 bp respectively. The results showed that SSR analysis produced a pattern that allowed a direct identification of cattle and sheep meat in raw and heat-treated samples. Also the results showed that SSR analysis provided with a rapid and effective method to detect the meat species and it could be easily identified and authenticated. In addition, the SSR PCR methods, as species-specific, are highly sensitive and will improve the detection limits for DNA sequences derived from these species. Also indirect application is the detection of disease cow (cattle) lunacy by PCR methods for authentication of cow (cattle) meat from origin country.

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.

scholarly journals ISOLASI FRAGMEN GEN PENYANDI PUTRESIN N-METILTRANSFERASE DAN QUINOLINAT FOSFORIBOSILTRANSFERASE ASAL TEMBAKAU LOKAL TEMANGGUNG (Nicotiana tabacum)

2020 ◽  
Vol 17 (3) ◽  
pp. 109 ◽  
Author(s):  
SESANTI BASUKI ◽  
NURHAJATI AA MATTJIK ◽  
SUWARSO SUWARSO ◽  
DESTA WIRNAS ◽  
SUDARSONO SUDARSONO
Keyword(s):  
Dna Sequences ◽  
Pcr Amplification ◽  
Data Bank ◽  
Degenerate Primer ◽  
Gene Encoding ◽  
Methyl Transferase ◽  
Dna Database ◽  
Genes Encoding ◽  
Tobacco Cultivar ◽  
Dna Template

<p>ABSTRAK</p><p>Upaya untuk menurunkan kandungan nikotin merupakan salah satuprioritas utama penelitian tembakau. Nikotin adalah senyawa alkaloidutama berpotensi dikonversi menjadi senyawa nor-nikotin yang bersifatkarsinogen. Gen PMT sebagai penyandi enzim putresin n-metiltransferase(PMT) dan gen QPT - penyandi enzim quinolinat fosforibosiltransferase(QPT) merupakan dua gen kunci yang berperan penting pada proses bio-sintesis nikotin. Penelitian ini bertujuan untuk mengisolasi potongan genPMT dan QPT asal tembakau lokal Indonesia, mengkarakterisasi danmenganalisis runutan DNA-nya. Tahapan penelitian dimulai dengan me-rancang primer degenerate berdasarkan informasi yang ada di pangkalandata Bank Gen NCBI (National Centre for Biotechnology Information),mengamplifikasi PCR menggunakan templat DNA genomik tembakaulokal cv. Sindoro1, mengklon potongan DNA hasil PCR dan menentukanrunutan DNA-nya. Hasil penelitian menunjukkan dari dua belas pasangprimer degenerate yang dirancang, hanya dua pasang primer yang meng-hasilkan potongan DNA hasil amplifikasi PCR, yaitu pasangan primerPMt-7 (F &amp; R) untuk gen PMT dan primer QPt-3 (F &amp; R) untuk gen QPT.Setelah dilakukan penentuan runutan DNA-nya, amplikon yang didapatdari hasil PCR dengan pasangan primer PMt-7 sebesar 1418 bp, sedangkanuntuk primer QPt-3 sebesar 205 bp. Runutan DNA gen PMT dan gen QPTasal tembakau lokal cv. Sindoro1 mempunyai tingkat kesamaan yang ting-gi dengan gen PMT dan gen QPT asal tembakau lainnya yang ada dipangkalan data Bank Gen NCBI.</p><p>Kata kunci : Gen PMT, gen QPT, lintasan biosintesis nikotin, perunutanDNA, amplifikasi PCR, primer degenerate</p><p>ABSTRACT</p><p>Isolation of Genes encoding Putrescine N-Methyl-transferase and Quinolinat Phosphoribosyl transferasederived from Temanggung Tobacco Cultivar (Nicotianatabacum)</p><p>Reduction of nicotine content is one of the major objective intobacco research. Nicotine is the main alcaloid compound that potentiallycould be converted into a carcinogenic compound (nor-nicotine). The PMTgene encoding putrescine N-methyl transferase (PMT) and the QPT gene -encoding quinolinate phosphoribosyl transferase (QPT) are the two keyenzymes involved in nicotine biosynthesis. The objectives of this researchwere to isolate PMT and QPT gene fragments originated from Indonesianlocal tobacco, to characterize, and to analyze their DNA sequences. Theresearch activities included: degenerate primer design based oninformation available in the GenBank DNA Database NCBI (NationalCentre for Biotechnology Information), PCR amplification usingdegenerate primer and genomic DNA template of a local tobacco cv.Sindoro1, clone the PCR amplified products, and determine their DNAnucleotide sequences. Results of the experiment indicated that from 12degenerate primer pairs synthesized, only two were able to yield positivePCR amplified products. These primer pairs were PMt-7 (F &amp; R primers)for PMT and QPt-3 (F &amp; R primers) for QPT. After DNA sequencing, theamplified DNA product amplified using PMt-7 degenerate primer pairswere 1418 bp, while that using QPt-3 primer pairs were only 205 bp.Nucleotide sequences of PMT or QPT gene fragments originated fromlocal tobacco cv. Sindoro1 showed a high nucleotide sequences identity ascompared to that of the respective genes from other tobacco species thatwere available in the GenBank DNA Database NCBI.</p><p>Key words: PMT gene, QPT gene, nicotine biosynthetic pathways, DNAsequencing, PCR amplification, degenerate primer</p>

PCR Identification of Beef, Sheep, Goat, and Pork in Raw and Heat-Treated Meat Mixtures

2004 ◽  
Vol 67 (1) ◽  
pp. 172-177 ◽  
Author(s):  
MIGUEL A. RODRÍGUEZ ◽  
TERESA GARCÍA ◽  
ISABEL GONZÁLEZ ◽  
LUIS ASENSIO ◽  
PABLO E. HERNÁNDEZ ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Bos Taurus ◽  
Ovis Aries ◽  
Capra Hircus ◽  
Accurate Identification ◽  
Qualitative Detection ◽  
Heat Treated ◽  
Sus Scrofa Domesticus ◽  
Species Specific ◽  
Identification Analysis

A PCR assay has been developed for the specific and qualitative detection of pork ( Sus scrofa domesticus), beef ( Bos taurus), sheep ( Ovis aries), and goat ( Capra hircus) in raw and heat-treated meat mixtures. A forward common primer was designed on a conserved DNA sequence in the mitochondrial 12S ribosomal RNA gene (rRNA), and reverse primers were designed to hybridize on species-specific DNA sequences of each species considered. The different sizes of the species-specific amplicons, separated by agarose gel electrophoresis, allowed clear species identification. Analysis of experimental meat mixtures demonstrated that the detection limit of the assay was 1% (wt/wt) for each species analyzed. This assay can be useful for the accurate identification of these species, avoiding mislabeling or fraudulent species substitution in meat mixtures.

Molecular identification of Helicobacter DNA present in human colorectal adenocarcinomas by 16S rDNA PCR amplification and pyrosequencing analysis

2005 ◽  
Vol 54 (11) ◽  
pp. 1031-1035 ◽  
Author(s):  
Niclas Grahn ◽  
Mounira Hmani-Aifa ◽  
Karin Fransén ◽  
Peter Söderkvist ◽  
Hans-Jürg Monstein
Keyword(s):  
16S Rdna ◽  
Dna Sequences ◽  
Pcr Amplification ◽  
Limited Information ◽  
Rdna Sequences ◽  
Biopsy Specimens ◽  
16S Rdna Pcr ◽  
H Pylori ◽  
Species Specific ◽  
Pyrosequencing Analysis

Seroepidemiological studies have indicated that Helicobacter pylori infection might be a possible risk factor for colorectal adenocarcinoma (CRC) development. However, limited information is available as to whether or not Helicobacter species are present in CRC tissues. In this study the presence of Helicobacter DNA in 77 CRC biopsies was investigated by means of a Helicobacter species-specific 16S rDNA PCR assay and real-time DNA pyrosequencing of the 16S rDNA variable V3 region. Pyrosequencing revealed the presence of Helicobacter DNA sequences in 21 of 77 biopsy specimens (27 %). 16S rDNA sequences corresponding to H. pylori 26695 and H. pylori J99 were most commonly found. Intriguingly, one sequence belonged to Helicobacter mustelae, previously identified in ferrets. No significant correlations were found in the prevalence of Helicobacter DNA between colon and rectum tumour biopsies (P = 0.815), nor between Dukes’ classes A/B and C/D (P = 0.262). 16S rDNA PCR amplification combined with pyrosequencing analysis of 16S rDNA variable V3 regions provides a powerful molecular tool to identify Helicobacter species in human biopsy specimens.

scholarly journals PolyCRACKER, a robust method for the unsupervised partitioning of polyploid subgenomes by signatures of repetitive DNA evolution

2018 ◽  
Author(s):  
Sean P. Gordon ◽  
Joshua J. Levy ◽  
John P. Vogel
Keyword(s):  
Repetitive Dna ◽  
Dna Sequences ◽  
Related Species ◽  
Software Package ◽  
Gc Content ◽  
Individual Species ◽  
Closely Related Species ◽  
As Species ◽  
Pooled Dna ◽  
Species Specific

AbstractExisting methods for assigning sequences to individual species from pooled DNA samples rely on differences in genome properties like GC content or sequences from related species. These approaches do not work for closely related species where gross features are indistinguishable and related genomes are lacking. We describe a method and associated software package that uses rapidly evolving repetitive DNA to circumvent these limitations. By using short, repetitive, DNA sequences as species-specific signals we separated closely related genomes without any prior knowledge. This approach is ideal for separating the subgenomes of polyploid species with unsequenced or unknown progenitor genomes.

Highly repeated DNA sequences as species-specific probes for Brugia

1987 ◽  
Vol 3 (12) ◽  
pp. 378-379 ◽  
Author(s):  
W.F. Piessens ◽  
L.A. McReynolds ◽  
S.A. Williams
Keyword(s):  
Dna Sequences ◽  
Repeated Dna ◽  
Repeated Dna Sequences ◽  
Highly Repeated Dna ◽  
As Species ◽  
Species Specific

scholarly journals Toxin Genes and Other Characteristics ofStaphylococcus aureus Isolates from Milk of Cows with Mastitis

2001 ◽  
Vol 8 (5) ◽  
pp. 959-964 ◽  
Author(s):  
Ö. Akineden ◽  
C. Annemüller ◽  
A. A. Hassan ◽  
C. Lämmler ◽  
W. Wolter ◽  
...  
Keyword(s):  
Protein A ◽  
Bovine Mastitis ◽  
Pcr Amplification ◽  
23S Rrna ◽  
Staphylococcal Enterotoxin A ◽  
Binding Region ◽  
Gene Encoding ◽  
Genes Encoding ◽  
Species Specific

ABSTRACT In the present study, 103 Staphylococcus aureusstrains isolated from milk samples from 60 cows with mastitis from eight different farms in seven different locations in one region of Germany were compared pheno- and genotypically and by identification of various toxins. On the basis of culture and hemolytic properties and by determination of the tube coagulase reaction, all of the isolates could be identified as S. aureus. This could be confirmed by PCR amplification of species-specific parts of the gene encoding the 23S rRNA. In addition, all of the S. aureus isolates harbored the genes encoding staphylococcal coagulase and clumping factor and the genes encoding the X region and the immunoglobulin G binding region of protein A. These four genes displayed size polymorphisms. By PCR amplification, the genes for the toxins staphylococcal enterotoxin A (SEA), SEC, SED, SEG, SEI, SEJ, and TSST-1 but not those for SEB, SEE, SEH, and the exfoliative toxins ETA and ETB could be detected. To analyze the epidemiological relationships, the isolates were subjected to DNA fingerprinting by macrorestriction analysis of their chromosomal DNAs. According to the observed gene polymorphisms, the toxin patterns, and the information given by macrorestriction analysis of the isolates by pulsed-field gel electrophoresis, a limited number of clones seemed to be responsible for the cases of bovine mastitis on the various farms.

scholarly journals DMSO Improves the Ski-Slope Effect in Direct PCR

2021 ◽  
Vol 11 (4) ◽  
pp. 1943
Author(s):  
Joo-Young Kim ◽  
Ju Yeon Jung ◽  
Da-Hye Kim ◽  
Seohyun Moon ◽  
Won-Hae Lee ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Pcr Amplification ◽  
Analytical Techniques ◽  
Direct Pcr ◽  
Dna Profile ◽  
Novel Technologies ◽  
Specific Amplification ◽  
Polymerase Chain ◽  
Dna Profiles

Analytical techniques such as DNA profiling are widely used in various fields, including forensic science, and novel technologies such as direct polymerase chain reaction (PCR) amplification are continuously being developed in order to acquire DNA profiles efficiently. However, non-specific amplification may occur depending on the quality of the crime scene evidence and amplification methods employed. In particular, the ski-slope effect observed in direct PCR amplification has led to inaccurate interpretations of the DNA profile results. In this study, we aimed to reduce the ski-slope effect by using dimethyl sulfoxide (DMSO) in direct PCR. We confirmed that DMSO (3.75%, v/v) increased the amplification yield of large-sized DNA sequences more than that of small-sized ones. Using 50 Korean buccal samples, we further demonstrated that DMSO reduced the ski-slope effect in direct PCR. These results suggest that the experimental method developed in this study is suitable for direct PCR and may help to successfully obtain DNA profiles from various types of evidence at crime scenes.

scholarly journals CaptureSeq: Hybridization-Based Enrichment of cpn60 Gene Fragments Reveals the Community Structures of Synthetic and Natural Microbial Ecosystems

2021 ◽  
Vol 9 (4) ◽  
pp. 816
Author(s):  
Matthew G. Links ◽  
Tim J. Dumonceaux ◽  
E. Luke McCarthy ◽  
Sean M. Hemmingsen ◽  
Edward Topp ◽  
...  
Keyword(s):  
Microbial Community ◽  
Microbial Communities ◽  
Dna Sequences ◽  
Pcr Amplification ◽  
Shotgun Sequencing ◽  
Natural Ecosystems ◽  
Gene Markers ◽  
Microbial Profile ◽  
Microbial Ecosystems ◽  
Pcr Targeting

Background. The molecular profiling of complex microbial communities has become the basis for examining the relationship between the microbiome composition, structure and metabolic functions of those communities. Microbial community structure can be partially assessed with “universal” PCR targeting taxonomic or functional gene markers. Increasingly, shotgun metagenomic DNA sequencing is providing more quantitative insight into microbiomes. However, both amplicon-based and shotgun sequencing approaches have shortcomings that limit the ability to study microbiome dynamics. Methods. We present a novel, amplicon-free, hybridization-based method (CaptureSeq) for profiling complex microbial communities using probes based on the chaperonin-60 gene. Molecular profiles of a commercially available synthetic microbial community standard were compared using CaptureSeq, whole metagenome sequencing, and 16S universal target amplification. Profiles were also generated for natural ecosystems including antibiotic-amended soils, manure storage tanks, and an agricultural reservoir. Results. The CaptureSeq method generated a microbial profile that encompassed all of the bacteria and eukaryotes in the panel with greater reproducibility and more accurate representation of high G/C content microorganisms compared to 16S amplification. In the natural ecosystems, CaptureSeq provided a much greater depth of coverage and sensitivity of detection compared to shotgun sequencing without prior selection. The resulting community profiles provided quantitatively reliable information about all three domains of life (Bacteria, Archaea, and Eukarya) in the different ecosystems. The applications of CaptureSeq will facilitate accurate studies of host-microbiome interactions for environmental, crop, animal and human health. Conclusions: cpn60-based hybridization enriched for taxonomically informative DNA sequences from complex mixtures. In synthetic and natural microbial ecosystems, CaptureSeq provided sequences from prokaryotes and eukaryotes simultaneously, with quantitatively reliable read abundances. CaptureSeq provides an alternative to PCR amplification of taxonomic markers with deep community coverage while minimizing amplification biases.

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