PCR-RFLP Analysis of Mitochondrial DNA: Differentiation of Fish Species

Wolf, Christian; Burgener, Matthias; Hübner, Philipp; Lüthy, Jürg

doi:10.1006/fstl.2000.0630

Cited by 98 publications

(84 citation statements)

References 23 publications

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“…Previous researchers identified 16 species of gadoid fish by PCR and DNA sequencing (Calo-Mata et al, 2003). PCR-RFLP has also been described as the useful technique for the differentiation between Salmon species (Russell et al, 2000), Flatfish (Sotelo et al, 2001), Sturgeon-caviar (Ludwig, Debus, & Jenneckens, 2002) and 23 Bony Fish over a wide range of species (Wolf, Burgener, Hübner, & Lüthy, 2000).…”

Section: Resultsmentioning

confidence: 99%

PCR-RFLP Analysis of D-Loop mtDNA in Indonesian Domestic Waterfowl

2017

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Phylogenetic or population genetic studies have been focused on finding genetic variations in mitochondrial DNA (mtDNA) because mutations in this region contained five times higher than other genetic materials. The aim of this research was to generate PCR-RFLP D-loop mtDNA profiles of some Indonesian domestic waterfowl. The samples were 35 local ducks, 5 muscovy duck, and 5 geese. These local ducks included on seven types of Javanese ducks in Central Java, i.e. Magelang duck, Peking duck, Pengging duck, Tegal Branjangan duck, Tegal Jarakan duck, Tegal Blorong duck, and Tegal Lemahan duck. PCR technique was used to amplify the D-loop genes, and RFLP analysis of the D-loop was performed with AluI and RsaI. The results show that muscovy duck has no restriction sites in D-loop gene. Moreover, goose has a restriction site of AluI in D-loop gene at 550 bp and 187 bp; and it has no restriction site of RsaI. The analysis of RsaI-RFLP in mtDNA D-loop region of ducks resulted in 1 restriction fragment pattern which can be applied in the identification of duck to differentiate it from goose and muscovy duck. PCR-RFLP analysis duck D-loop mtDNA using AluI and RsaI restriction enzyme resulted in 3 combinations of restriction fragment pattern shown in 3 haplotypes (A, B, and C). Genetic diversity of Indonesian domestic waterfowl population could be identified by using PCR-RFLP analysis on mtDNA D-loop region. So far, there are few results of research on Indonesian domestic waterfowl genetic characteristics based on PCR-RFLP mtDNA D-loop marker. Also, this research has a good contribution on genetic characterization of local duck. The characterization is an important aspect to maintain breed quality and manage the genetic resources of Indonesian germplasm and in facilitating the breeding program.

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Section: Resultsmentioning

confidence: 99%

PCR-RFLP Analysis of D-Loop mtDNA in Indonesian Domestic Waterfowl

2017

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“…The optimal condition of pH, ionic strength, buffer concentration and temperatures were at pH 7.0, 1.0 M NaCl, 0.05 M of Na-phosphate buffer and 25 °C, respectively. The linear range of different concentration complementary DNA was within the range between 1.0 x 10 -13 M to 1.0 x 10 -8 M. This study was first reported the used of [Ru(bpy) 2 PIP] 2+ for detection of porcine oligonucleotides…”

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confidence: 81%

“…Kesan masa pemegunan dan penghibridan DNA Kajian kesan masa pemegunan DNA prob dilakukan pada sela masa 1 jam sehingga 24 jam manakala kajian terhadap masa penghibridan DNA sasaran dilakukan dengan pelbagai masa penghibridan antara 0.5 hingga 3.0 jam. Pemegunan 5.0 µM DNA prob dilakukan dalam larutan penimbal K-fosfat 0.05M (pH 7.0) manakala penghibridan dengan 2.0 µM DNA sasaran di dalam 0.05 M penimbal Na-fosfat (pH 7.0) yang juga mengandungi 30 µM larutan penunjuk redoks [Ru(bpy) 2 dengan DNA sasaran ditentukan dengan melakukan pengukuran arus puncak secara berturutan dengan menggunakan lima SPE yang sama keadaan. SPE yang telah terubah suai itu mengandungi kuantiti AuNPs, mikrosfera akrilik, kepekatan prob dan DNA sasaran, masa pemegunan dan penghibridan DNA serta kepekatan larutan penimbal fosfat dan kekuatan ion Na + Keselektifan biosensor DNA pada DNA sasaran Kajian keselektifan biosensor DNA sasaran porsin diuji dengan menggunakan jenis -jenis DNA sasaran yang berbeza iaitu DNA gallus (ayam), DNA bos taurus (daging lembu) dan DNA bukan sasaran porsin manakala DNA prob porsin dijadikan sebagai pengosong.…”

Section: Bahan Dan Kaedah Bahan Kimiaunclassified

“…The concentration of [Ru(bpy) 2 PIP] 2+ complex and DNA probe were found to be optimal at 50 µM and 2 µM, respectively. The optimal time for DNA probe immobilization and hybridization were 7 hours and 60 minutes, accordingly.…”

mentioning

confidence: 99%

“…DNA membawa maklumat genetik organisma dan maklumat yang wujud dalam DNA adalah lebih banyak daripada protein disebabkan oleh kemerosotan kod genetik yang berlaku apabila DNA mengalami perubahan bentuk daripada DNA kepada protein [2]. Pada masa kini, pengesanan DNA adalah satu bidang yang menarik minat penyelidik kerana aplikasinya yang meluas dalam bidang klinikal, forensik, farmaseutikal dan pemprosesan makanan [3].…”

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Fabrication of Porcine DNA Biosensor Based on Ruthenium Bipyridine Complex

Halid

Zakariah

Lee³

et al. 2016

MJAS

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AbstrakBiosensor DNA elektrokimia bagi pengesanan jujukan DNA porsin berasaskan kompleks rutenium (II) sebagai label aktif redoks telah dibangunkan. Sistem biosensor difabrikasi berdasarkan elektrod bercetak skrin (SPE) terubahsuai dengan zarah nano emas (AuNPs) sebagai transduser yang dipegunkan bersama mikrosfera poli (n-butilakrilat-N-akriloksisuksnimida) dan jujukan DNA prob terikat melalui ikatan kovalen. Kompleks interkalator rutenium (II), [Ru(bpy) 2+ dan DNA prob, masing-masing adalah 50 µM dan 2 µM. Masa yang optimum bagi pemegunan DNA prob dan penghibridan DNA sasaran adalah 7 jam dan 60 minit. Keadaan optimum bagi pH, kekuatan ion, kepekatan larutan penimbal dan suhu, masing-masing adalah pH 7.0, 1.0 M NaCl, 0.05 M potassium fosfat pada suhu 25°C. Julat linear bagi kepekatan DNA sasaran yang berbeza adalah antara 1.0 x 10 -13 M to 1.0 x 10 -8 M. Kajian ini adalah yang pertama melaporkan tentang penggunaan [Ru(bpy) 2 PIP] 2+ bagi pengesanan DNA porsin Kata kunci: biosensor, DNA porsin, kompleks rutenium bipiridina Abstract Electrochemical DNA biosensor for detection of porcine oligonucleotides based on ruthenium (II) as label redox complex has been developed. The system of biosensor is based on the modified screen printed carbon electrode (SPE) with goldnanoparticles (AuNPs) as transducer immobilized with poly(n-butylacrylate-N-acryloxysuccinimide) microsphere and porcine DNA probe sequences was attached onto it via the covalent bond. The ruthenium (

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Requirements for the application of protein sodium dodecyl sulfate-polyacrylamide gel electrophoresis and randomly amplified polymorphic DNA analyses to product speciation

2001

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Raw, cooked, fried, smoked and gravad (brine-cured) products were analyzed by Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of proteins and by randomly amplified polymorphic DNA (RAPD) in order to identify the species used in their manufacture. The discriminatory power of SDS-PAGE was dependent primarily on the composition and secondarily on the size of the gels: the Laemmli buffer system with 15% acrylamide and 0.087% piperazine diacrylamide separating gels resolved more discriminant protein bands than any of the commercial gels tested. Some of the processing conditions induced alterations in the protein patterns that made identification dubious. Differentiation even between closely related species was easier by RAPD than by SDS-PAGE. Neither the processing conditions nor the tissue from which the DNA was extracted had a significant effect on the RAPD profiles. For identifications based on SDS-PAGE, one should use an optimized gel composition and separate the sample under analysis in the same gel as the references. For RAPD-based identifications, the unknown sample should be amplified together with reference samples and separated in the same gel.

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PCR-RFLP Analysis of Mitochondrial DNA: Differentiation of Fish Species

Cited by 98 publications

References 23 publications

PCR-RFLP Analysis of D-Loop mtDNA in Indonesian Domestic Waterfowl

PCR-RFLP Analysis of D-Loop mtDNA in Indonesian Domestic Waterfowl

Fabrication of Porcine DNA Biosensor Based on Ruthenium Bipyridine Complex

Requirements for the application of protein sodium dodecyl sulfate-polyacrylamide gel electrophoresis and randomly amplified polymorphic DNA analyses to product speciation

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