A Simple, Inexpensive and Safe Method for DNA Extraction of Frigid and Clotted Blood Samples
Novelty in Biomedicine,
Vol. 3 No. 3 (2015),
22 July 2015
,
Page 119-123
https://doi.org/10.22037/nbm.v3i3.9507
Abstract
Background: Extraction of blood genomicDNAis one of the main approaches for clinical and molecular biology studies. Although several methods have been developed for extraction of blood genomic DNA, most of these methods consume long time and use expensive chemicals such as proteinase K and toxic organic solvent such as phenol and chloroform. The objective of this study was to developed easy and safe method forDNAextraction from clotted and frozen whole blood. This method has many advantages: time reducing, using inexpensive materials, without phenol and chloroform, achieving of high molecular weight and good quality genomicDNA.Materials and Methods: DNA extraction was performed by two methods (new and phenol-chloroform method). Then quantity and quality parameters were evaluated by 1% agarose gel electrophoresis, Nano drop analysis and efficiency of Polymerase Chain Reaction (PCR).
Results: Extracted DNA from 500μL of blood samples were 457.7ng/μl and 212ng/μL and their purity (OD260/OD280) were 1.8 and 1.81 for new recommended and phenol–chloroform methods respectively. The PCR results indicated that D16S539 and CSF1PO loci were amplified.
Conclusion: These results shown that this method is simple, fast, safe and most economical.
- Genomic DNA extraction
- Quality and quantity of DNA
- Blood sample
How to Cite
References
Samadi-Shams S, Zununi-Vahed S, Soltanzad F, Kafil V, Barzegari A, Atashpaz S, et al. Highly Effective DNA Extraction Method from Fresh, Frozen, Dried and Clotted Blood Samples. Bio Impacts. 2011;1(3):183-7.
Kalmar T, Bachrati CZ, Marcsik A, Rasko I. A Simple and efficient method for PCR amplifiable DNA extraction from ancient bones. Nucleic Acids Res. 2000;28:67-70.
Garg UC, Hanson NQ, Tsai MY, Ethfeldt JH. Simple and rapid method for extraction of DNA from fresh and cryopreserved clotted human blood. Clin Chem. 1996;42:647-8.
Leland JC, Kirakosyan A, Kaufman PB, Westfall MV (Editors). Handbook of Molecular and Cellular Methods in Biology and Medicine, 3th ed. CRC Press, Taylor and forensic group, United States of America. 2011;1-146.
Loffler J, Hebart H, Schumacher U, Reitze H, Einsele H. Comparison of different methods for extraction of DNA of fungal pathogens from cultures and blood. Clin Microbial. 1997;35:3311-2.
Barbaro A, Staiti N, Cormaci P Saravo L. DNA profiling by different extraction methods. International Congress Series, Elsevier. 2004;1261:562–4.
Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, New York. 1989.
Dokanehiifard S, Bidmeshkipour A. Comparison of three methods for Hepatitis B Virus DNA extraction from infected serum samples. EJBS. 2010;2(1):26-30.
Radheshyam M, Brijesh K, Shyam S. Evaluation of salt-out method for the isolation of DNA from whole blood: A pathological approach of DNA based diagnosis. IJLBPR. 2013;2(2):57-62.
Bank S, Andersen Nexø B, Andersen V, Vogel U, Andersen PS. High-Quality and -Quantity DNA Extraction from Frozen Archival Blood Clots for Genotyping of Single-Nucleotide Polymorphisms. Genet Test Mol Biomarkers. 2013;17(6):501-3.
Adeli K, Ogbonna G. Rapid purification of human DNA from whole blood for potential application in clinical chemistry laboratories. Clin Chem. 1990;36 (2):261-4.
Montgomery GW, Sise JA. Extraction of DNA From sheep white blood cells. Agr Res. 1990;33:437-441.
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