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Comparison of six simple methods for extracting ribosomal and mitochondrial DNA from Toxocara and Toxascaris nematodes

Mikaeili, F. and Kia, E.B. and Sharbatkhori, M. and Sharifdini, M. and Jalalizand, N. and Heidari, Z. and Zarei, Z. and Stensvold, C.R. and Mirhendi, H. (2013) Comparison of six simple methods for extracting ribosomal and mitochondrial DNA from Toxocara and Toxascaris nematodes. Experimental Parasitology, 134 (2). pp. 155-159. ISSN 00144894 (ISSN)

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Abstract

Six simple methods for extraction of ribosomal and mitochondrial DNA from Toxocara canis, Toxocara cati and Toxascaris leonina were compared by evaluating the presence, appearance and intensity of PCR products visualized on agarose gels and amplified from DNA extracted by each of the methods. For each species, two isolates were obtained from the intestines of their respective hosts: T. canis and T. leonina from dogs, and T. cati from cats. For all isolates, total DNA was extracted using six different methods, including grinding, boiling, crushing, beating, freeze-thawing and the use of a commercial kit. To evaluate the efficacy of each method, the internal transcribed spacer (ITS) region and the cytochrome c oxidase subunit 1 (cox1) gene were chosen as representative markers for ribosomal and mitochondrial DNA, respectively. Among the six DNA extraction methods, the beating method was the most cost effective for all three species, followed by the commercial kit. Both methods produced high intensity bands on agarose gels and were characterized by no or minimal smear formation, depending on gene target; however, beating was less expensive. We therefore recommend the beating method for studies where costs need to be kept at low levels. © 2013 Elsevier Inc.

Item Type: Article
Additional Information: Unmapped bibliographic data: LA - English [Field not mapped to EPrints] J2 - Exp. Parasitol. [Field not mapped to EPrints] C2 - 23499880 [Field not mapped to EPrints] AD - Dept. of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran [Field not mapped to EPrints] AD - Dept. of Medical Parasitology and Mycology, School of Medicine, Golestan University of Medical Sciences, Gorgan, Iran [Field not mapped to EPrints] AD - National Institute of Health Research, Tehran University of Medical Sciences, Tehran, Iran [Field not mapped to EPrints] AD - Dept. of Microbiology and Infection control, Statens Serum Institut, Copenhagen, Denmark [Field not mapped to EPrints] DB - Scopus [Field not mapped to EPrints]
Uncontrolled Keywords: DNA extraction, Nematode, PCR, Toxascaris, Toxocara, agarose, cytochrome c oxidase, internal transcribed spacer, mitochondrial DNA, ribosome DNA, analytic method, analytical equipment, article, beating, boiling point, cat, controlled study, cost effectiveness analysis, cox1 gene, crushing, DNA extraction, dog, freeze thawing, gene amplification, grinding, intermethod comparison, intestine, nonhuman, polymerase chain reaction, priority journal, Toxascaris, Toxascaris leonina, Toxocara, Toxocara canis, Toxocara cati, Animals, Cats, DNA, Helminth, DNA, Mitochondrial, DNA, Ribosomal, Dogs, Electrophoresis, Agar Gel, Polymerase Chain Reaction, Toxascaris, Toxocara, Toxocara canis, Canis familiaris, Nematoda, Toxascaris, Toxascaris leonina, Toxocara, Toxocara canis, Toxocara cati
Subjects: مقالات نمایه شده محققین دانشگاه در سایت ,Web of Science ,Scopus
موارد کلی
Divisions: معاونت تحقیقات و فناوری
Depositing User: GOUMS
Date Deposited: 18 Apr 2015 04:14
Last Modified: 10 May 2015 09:17
URI: http://eprints.goums.ac.ir/id/eprint/1872

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