Effects of storage temperature on the quantity and integrity of genomic DNA extracted from mice tissues: A comparison of recovery methods

Efficient extraction of genomic DNA (gDNA) from biological materials found in harsh environments is the first step for successful forensic DNA profiling. This study aimed to evaluate two methods for DNA recovery from animal tissues (livers, muscles), focusing on the best storage temperature for DNA...

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Main Authors: Huda H. Al-Griw, Zena A. Zraba, Salsabiel K. Al-Muntaser, Marwan M. Draid, Aisha M. Zaidi, Refaat M. Tabagh, Mohamed A. Al-Griw
Format: Article
Language:English
Published: Tripoli University 2017-08-01
Series:Open Veterinary Journal
Subjects:
Online Access:http://www.openveterinaryjournal.com/OVJ-2017-05-090%20H.H.%20Al-Griw%20et%20al.pdf
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spelling doaj-0fccc6b663a344c3988ae4d1ca8532602021-10-02T03:57:43ZengTripoli UniversityOpen Veterinary Journal2218-60502218-60502017-08-0173239243http://dx.doi.org/10.4314/ovj.v7i3.7Effects of storage temperature on the quantity and integrity of genomic DNA extracted from mice tissues: A comparison of recovery methodsHuda H. Al-Griw0Zena A. Zraba1Salsabiel K. Al-Muntaser2Marwan M. Draid3Aisha M. Zaidi4Refaat M. Tabagh5Mohamed A. Al-Griw6Department of Microbiology and Parasitology, Faculty of Veterinary Medicine, University of Tripoli, Tripoli, LibyaDepartment of Forensic Biology, Faculty of Science, University of Tripoli. Tripoli, LibyaDepartment of Forensic Biology, Faculty of Science, University of Tripoli. Tripoli, LibyaDepartment of Pharmacology, Toxicology and Forensic Medicine, Faculty of Veterinary Medicine, University of Tripoli, Tripoli, LibyaDepartment of Physiology, Biochemistry and Animal Nutrition, Faculty of Veterinary Medicine, University of Tripoli, Tripoli, LibyaDepartment of Forensic Biology, Faculty of Science, University of Tripoli. Tripoli, LibyaDepartment of Forensic Biology, Faculty of Science, University of Tripoli. Tripoli, LibyaEfficient extraction of genomic DNA (gDNA) from biological materials found in harsh environments is the first step for successful forensic DNA profiling. This study aimed to evaluate two methods for DNA recovery from animal tissues (livers, muscles), focusing on the best storage temperature for DNA yield in term of quality, quantity, and integrity for use in several downstream molecular techniques. Six male Swiss albino mice were sacrificed, liver and muscle tissues (n=32) were then harvested and stored for one week in different temperatures, -20C, 4C, 25C and 40C. The conditioned animal tissues were used for DNA extraction by Chelex-100 method or NucleoSpin Blood and Tissue kit. The extracted gDNA was visualized on 1.5% agarose gel electrophoresis to determine the quality of gDNA and analysed spectrophotometrically to determine the DNA concentration and the purity. Both methods, Chelex-100 and NucleoSpin Blood and Tissue kit found to be appropriate for yielding high quantity of gDNA, with the Chelex100 method yielding a greater quantity (P < 0.045) than the kit. At -20C, 4C, and 25C temperatures, the concentration of DNA yield was numerically lower than at 40C. The NucleoSpin Blood and Tissue kit produced a higher (P=0.031) purity product than the Chelex-100 method, particularly for muscle tissues. The Chelex-100 method is cheap, fast, effective, and is a crucial tool for yielding DNA from animal tissues (livers, muscles) exposed to harsh environment with little limitations.http://www.openveterinaryjournal.com/OVJ-2017-05-090%20H.H.%20Al-Griw%20et%20al.pdfDNA degradationDNA extractionDNA profilingPurityTemperature
collection DOAJ
language English
format Article
sources DOAJ
author Huda H. Al-Griw
Zena A. Zraba
Salsabiel K. Al-Muntaser
Marwan M. Draid
Aisha M. Zaidi
Refaat M. Tabagh
Mohamed A. Al-Griw
spellingShingle Huda H. Al-Griw
Zena A. Zraba
Salsabiel K. Al-Muntaser
Marwan M. Draid
Aisha M. Zaidi
Refaat M. Tabagh
Mohamed A. Al-Griw
Effects of storage temperature on the quantity and integrity of genomic DNA extracted from mice tissues: A comparison of recovery methods
Open Veterinary Journal
DNA degradation
DNA extraction
DNA profiling
Purity
Temperature
author_facet Huda H. Al-Griw
Zena A. Zraba
Salsabiel K. Al-Muntaser
Marwan M. Draid
Aisha M. Zaidi
Refaat M. Tabagh
Mohamed A. Al-Griw
author_sort Huda H. Al-Griw
title Effects of storage temperature on the quantity and integrity of genomic DNA extracted from mice tissues: A comparison of recovery methods
title_short Effects of storage temperature on the quantity and integrity of genomic DNA extracted from mice tissues: A comparison of recovery methods
title_full Effects of storage temperature on the quantity and integrity of genomic DNA extracted from mice tissues: A comparison of recovery methods
title_fullStr Effects of storage temperature on the quantity and integrity of genomic DNA extracted from mice tissues: A comparison of recovery methods
title_full_unstemmed Effects of storage temperature on the quantity and integrity of genomic DNA extracted from mice tissues: A comparison of recovery methods
title_sort effects of storage temperature on the quantity and integrity of genomic dna extracted from mice tissues: a comparison of recovery methods
publisher Tripoli University
series Open Veterinary Journal
issn 2218-6050
2218-6050
publishDate 2017-08-01
description Efficient extraction of genomic DNA (gDNA) from biological materials found in harsh environments is the first step for successful forensic DNA profiling. This study aimed to evaluate two methods for DNA recovery from animal tissues (livers, muscles), focusing on the best storage temperature for DNA yield in term of quality, quantity, and integrity for use in several downstream molecular techniques. Six male Swiss albino mice were sacrificed, liver and muscle tissues (n=32) were then harvested and stored for one week in different temperatures, -20C, 4C, 25C and 40C. The conditioned animal tissues were used for DNA extraction by Chelex-100 method or NucleoSpin Blood and Tissue kit. The extracted gDNA was visualized on 1.5% agarose gel electrophoresis to determine the quality of gDNA and analysed spectrophotometrically to determine the DNA concentration and the purity. Both methods, Chelex-100 and NucleoSpin Blood and Tissue kit found to be appropriate for yielding high quantity of gDNA, with the Chelex100 method yielding a greater quantity (P < 0.045) than the kit. At -20C, 4C, and 25C temperatures, the concentration of DNA yield was numerically lower than at 40C. The NucleoSpin Blood and Tissue kit produced a higher (P=0.031) purity product than the Chelex-100 method, particularly for muscle tissues. The Chelex-100 method is cheap, fast, effective, and is a crucial tool for yielding DNA from animal tissues (livers, muscles) exposed to harsh environment with little limitations.
topic DNA degradation
DNA extraction
DNA profiling
Purity
Temperature
url http://www.openveterinaryjournal.com/OVJ-2017-05-090%20H.H.%20Al-Griw%20et%20al.pdf
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