Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Al-Griw, Huda H., Zraba, Zena A., Al-Muntaser, Salsabiel K., Draid, Marwan M., Zaidi, Aisha M., Tabagh, Refaat M., Al-Griw, Mohamed A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Faculty of Veterinary Medicine, University of Tripoli and Libyan Authority for Research, Science and Technology 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5579564/
https://www.ncbi.nlm.nih.gov/pubmed/28884076
http://dx.doi.org/10.4314/ovj.v7i3.7
_version_ 1783260730389168128
author Al-Griw, Huda H.
Zraba, Zena A.
Al-Muntaser, Salsabiel K.
Draid, Marwan M.
Zaidi, Aisha M.
Tabagh, Refaat M.
Al-Griw, Mohamed A.
author_facet Al-Griw, Huda H.
Zraba, Zena A.
Al-Muntaser, Salsabiel K.
Draid, Marwan M.
Zaidi, Aisha M.
Tabagh, Refaat M.
Al-Griw, Mohamed A.
author_sort Al-Griw, Huda H.
collection PubMed
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 NucleoSpinC 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 NucleoSpinC 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.
format Online
Article
Text
id pubmed-5579564
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher Faculty of Veterinary Medicine, University of Tripoli and Libyan Authority for Research, Science and Technology
record_format MEDLINE/PubMed
spelling pubmed-55795642017-09-07 Effects of storage temperature on the quantity and integrity of genomic DNA extracted from mice tissues: A comparison of recovery methods Al-Griw, Huda H. Zraba, Zena A. Al-Muntaser, Salsabiel K. Draid, Marwan M. Zaidi, Aisha M. Tabagh, Refaat M. Al-Griw, Mohamed A. Open Vet J Original Article 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 NucleoSpinC 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 NucleoSpinC 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. Faculty of Veterinary Medicine, University of Tripoli and Libyan Authority for Research, Science and Technology 2017 2017-08-05 /pmc/articles/PMC5579564/ /pubmed/28884076 http://dx.doi.org/10.4314/ovj.v7i3.7 Text en Copyright: © Open Veterinary Journal http://creativecommons.org/licenses/by-nc-sa/4.0 Open Veterinary Journal is licensed under a Creative Commons Attribution 4.0 International License.
spellingShingle Original Article
Al-Griw, Huda H.
Zraba, Zena A.
Al-Muntaser, Salsabiel K.
Draid, Marwan M.
Zaidi, Aisha M.
Tabagh, Refaat M.
Al-Griw, Mohamed A.
Effects of storage temperature on the quantity and integrity of genomic DNA extracted from mice tissues: A comparison of recovery methods
title 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_short 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
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5579564/
https://www.ncbi.nlm.nih.gov/pubmed/28884076
http://dx.doi.org/10.4314/ovj.v7i3.7
work_keys_str_mv AT algriwhudah effectsofstoragetemperatureonthequantityandintegrityofgenomicdnaextractedfrommicetissuesacomparisonofrecoverymethods
AT zrabazenaa effectsofstoragetemperatureonthequantityandintegrityofgenomicdnaextractedfrommicetissuesacomparisonofrecoverymethods
AT almuntasersalsabielk effectsofstoragetemperatureonthequantityandintegrityofgenomicdnaextractedfrommicetissuesacomparisonofrecoverymethods
AT draidmarwanm effectsofstoragetemperatureonthequantityandintegrityofgenomicdnaextractedfrommicetissuesacomparisonofrecoverymethods
AT zaidiaisham effectsofstoragetemperatureonthequantityandintegrityofgenomicdnaextractedfrommicetissuesacomparisonofrecoverymethods
AT tabaghrefaatm effectsofstoragetemperatureonthequantityandintegrityofgenomicdnaextractedfrommicetissuesacomparisonofrecoverymethods
AT algriwmohameda effectsofstoragetemperatureonthequantityandintegrityofgenomicdnaextractedfrommicetissuesacomparisonofrecoverymethods