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Influence of Storage Methods of Whole Blood Samples on DNA Integrity in Epidemiological Studies
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Objectives: To investigate the influence of storage methods of whole blood samples on Deoxyribonucleic acid (DNA) quality.
Method and materials: Whole blood samples were collected from 90 patients reported to the Government Tertiary-care Dental College and Hospital, Tamil Nadu, India. The blood samples were stored at three different temperatures -80 °C, -20 °C, 4 °C which were analyzed for DNA quality immediately within 4 h and after a storage period of 60 days. Quantification of DNA in the sample was recorded using a spectrophotometer..
Results: The average concentration of isolated DNA ranged from 0.5 ng/μl to 154.2 ng/μl with purity A260/A280ratio ranging from 0.7 to 2.0. The mean DNA concentration H = 97.2 (p = 0.001) and purity H = 80.1 (p = 0.001) obtained from whole blood stored at 4 °C were statistically significant. Thus samples stored at 4 °C did not affect on DNA yield and quality..
Conclusion: We conclude that storage at 4 8C temperature for whole blood does not substantially affect the molecular assay measurements. This study enlightens the influence of storage temperatures and duration in genetic epidemiology.
Method and materials: Whole blood samples were collected from 90 patients reported to the Government Tertiary-care Dental College and Hospital, Tamil Nadu, India. The blood samples were stored at three different temperatures -80 °C, -20 °C, 4 °C which were analyzed for DNA quality immediately within 4 h and after a storage period of 60 days. Quantification of DNA in the sample was recorded using a spectrophotometer..
Results: The average concentration of isolated DNA ranged from 0.5 ng/μl to 154.2 ng/μl with purity A260/A280ratio ranging from 0.7 to 2.0. The mean DNA concentration H = 97.2 (p = 0.001) and purity H = 80.1 (p = 0.001) obtained from whole blood stored at 4 °C were statistically significant. Thus samples stored at 4 °C did not affect on DNA yield and quality..
Conclusion: We conclude that storage at 4 8C temperature for whole blood does not substantially affect the molecular assay measurements. This study enlightens the influence of storage temperatures and duration in genetic epidemiology.
Keywords
DNA Isolation, Genetic Epidemiology, Whole Blood.
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- Johnson L, Genco RJ, Damsky C, et al. Genetics and its implications for clinical dental practice and education: report of panel 3 of the Macy study. J Dent Educ. 2008;72 (suppl):86–94.
- Venter JC, Adams MD, Myers EW, et al. The sequence of the human genome. Science. 2001;29:1304–1351.
- Richardson AJ, Narendran N, Guymer RH, Vu H, Baird PN. Blood storage at 4 8C-factors involved in DNA yield and quality. J Lab Clin Med. 2006;147:290–294.
- Cushwa WT, Medrano JF. Effects of blood storage time and temperature on DNA yield and quality. Biotechniques. 1993;14:204–207.
- Visvikis S, Schlenck A, Maurice M. DNA extraction and stability for epidemiological studies. Clin Chem Lab Med. 1998;36:551–555.
- Suuriniemi M, Mahonen A, Kovanen V, Alen MCS. Relation of PvuII site polymorphism in the COL1A2 gene to the risk of fractures in prepubertal Finnish girls. Physiol Genomics. 2003;14:217–224.
- Mychaleckyj JC, Farber EA, Chmielewski J, Artale J, Light LS, Bowden DW. Buffy coat specimens remain viable as a DNA source for highly multiplexed genome-wide genetic tests after long term storage. J Transl Med BioMed Central Ltd. 2011;9:91.
- Carpi FM, Di Pietro F, Vincenzetti S, Mignini F, Napolioni V. Human DNA extraction methods: patents and applications. Recent Pat DNA Gene Seq. 2011;5:1–7.
- Dickinson JL, Sale MM, Craig JE, Mackey DA. Laboratory methods in ophthalmic genetics: obtaining DNA from patients. Ophthalmic Genet. 2001;22:49–60.
- Abraham JE, Maranian MJ, Spiteri I, et al. Saliva samples are a viable alternative to blood samples as a source of DNA for high throughput genotyping. BMC Med Genomics. 2012;5:1–6.
- Philibert Ra, Zadorozhnyaya O, Beach SRH, Brody GH. Comparison of the genotyping results using DNA obtained from blood and saliva. Psychiatr Genet. 2008;18:275–281.
- Zhou J, Wu J, Zhao X, et al. Biostorage and quality control for human peripheral blood leukocytes. Biopreserv Biobank. 2015;13:13–19.
- Lahiri DK, Schnabel B. DNA isolation by a rapid method from human blood samples: effects of MgCl2, EDTA, storage time and temperature on DNA yield and quality. Biochem Genet. 1993;31:321–328.
- Madisen L, Hoar DI, Holroyd CD, Crisp M, Hodes ME. DNA banking: the effects of storage of blood and isolated DNA on the integrity of DNA. Am J Med Genet. 1987;27:379–390.
- Howlett SE, Castillo HS, Gioeni LJ, et al. Evaluation of DNA stable for DNA storage at ambient temperature. Forensic Sci Int Genet. 2014;8:170–178.
- Hansen TV, Simonsen MK, Nielsen FC, Hundrup YA. Collection of blood, saliva, and buccal cell samples in a pilot study on the Danish nurse cohort: comparison of the response rate and quality of genomic DNA. Cancer Epidemiol Biomarkers Prev. 2007;16:2072–2076.
- Udtha M, Flores R, Sanner J, et al. The protection and stabilization of whole blood at room temperature. Biopreserv Biobank. 2014;12:332–336.
- Rogers NL, Cole SA, Lan HC, Crossa A, Demerath EW. New saliva DNA collection method compared to buccal cell collection techniques for epidemiological studies. Am J Hum Biol. 2007;19:319–326.
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