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. 2017;7(3):239-243.
doi: 10.4314/ovj.v7i3.7. Epub 2017 Aug 5.

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

Huda H Al-Griw  1 , Zena A Zraba  2 , Salsabiel K Al-Muntaser  2 , Marwan M Draid  3 , Aisha M Zaidi  4 , Refaat M Tabagh  2 , Mohamed A Al-Griw  2
Affiliations

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

Huda H Al-Griw et al. Open Vet J. 2017.

Abstract

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.

Keywords: DNA degradation; DNA extraction; DNA profiling; Purity; Temperature.

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Figures

Fig. 1
Fig. 1
Average concentration of DNA measured in ng/µl. DNA was extracted from animal tissues (muscles, livers) stored in different temperature conditions using NucleoSpinl Blood & Tissue kit (black bar) and Chelex-100 method (white bar). Data are expressed as mean ± S.D, P < 0.05 significant.
Fig. 2
Fig. 2
Average purity of extracted gDNA. DNA was spectrophotometrically analysed to determine the purity (A260/A280) of extracted DNA from animal tissues (muscles, livers) stored in different temperature conditions using NucleoSpinl Blood & Tissue kit (black bar) and Chelex-100 method (white bar). Data are expressed as mean ± S.D, P < 0.05 significant.
Fig. 3
Fig. 3
Agarose gel electrophoresis results on 1% agarose gel with DNA extracted from four animal tissue samples (2-5) by using the Chelex-100 method. Lanes: 1 and 6 ladders; lanes 2-5 liver DNA samples stored in different temperatures, L4°C, L 25°C, L-20°C and L 40°C respectively.
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