Effect Of Blood Storage Conditions On The Integrity Of  Human Dna

Jaya Tiwari; Vijaylakshmi Jain; Meenakshi Mishra; Rajkiran Sahu; Azima Khushnud; P K Khodiar; Pankaj Kishor Mishra

doi:10.48165/

Authors

Jaya Tiwari Division of Medical Biotechnology, Department of Biochemistry, Pt. Jawahar Lal Nehru Memorial Medical College, Raipur - 492 001, Chhattisgarh (India)
Vijaylakshmi Jain Department of Molecular & Cellular Engineering, Sam Higginbottom University of Agriculture, Technology & Sciences, Allahabad - 211 007, Uttar Pradesh (India)
Meenakshi Mishra School of Life and Allied Sciences, ITM University Atal Nagar Raipur - 493 661 (India)
Rajkiran Sahu Division of Medical Biotechnology, Department of Biochemistry, Pt. Jawahar Lal Nehru Memorial Medical College, Raipur - 492 001, Chhattisgarh (India)
Azima Khushnud Division of Medical Biotechnology, Department of Biochemistry, Pt. Jawahar Lal Nehru Memorial Medical College, Raipur - 492 001, Chhattisgarh (India)
P K Khodiar Division of Medical Biotechnology, Department of Biochemistry, Pt. Jawahar Lal Nehru Memorial Medical College, Raipur - 492 001, Chhattisgarh (India)
Pankaj Kishor Mishra Division of Medical Biotechnology, Department of Biochemistry, Pt. Jawahar Lal Nehru Memorial Medical College, Raipur - 492 001, Chhattisgarh (India)

DOI:

https://doi.org/10.48165/

Keywords:

CYP1A1, DNA analysis, PCR, phenol-chloroform, salting out

Abstract

A study was performed in blood samples, collected from 15 healthy individuals during July to December 2017, to evaluate the integrity of DNA by preserving blood samples at three temperatures i.e. 4, -20 and -80ºC as against the fresh sample (control) for an incubation period of 30 days. DNA was extracted by standard protocol of phenol-chloroform and salting out method. The effect of temperature on DNA quality and quantity was analyzed spectrophotometerically, followed by amplification of Cytochrome p450 1A1 (CYP1A1) gene. In this prognosis, the study revealed that higher purity was attained at 4ºC, yield and concentration at -20ºC on extraction by phenol-chloroform. On contrary, in salting out method maximal yield and concentration was acquired at 4ºC and purity at -20ºC. Comparison of methods showed that phenol-chloroform method gave best result. DNA obtained from preserved blood samples is appropriate for amplification and further studies.

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References

AIRokayan, S.A.H. 2000. Effect of storage temperature on the quality and quantity of DNA extracted from blood. Pakistan Journal of Biological Sciences, 3: 392-394.

Albirno, C.G. and Romanowski, V. 1994. Phenol extraction revisited a rapid method for the isolation and preservation of human genomic DNA from whole blood. Molecular and Cellular Probes, 8: 423-427.

Bulla, A., Witt, B.D., Ammerlaan, W., Betsou, F. and Lescuyer, P. 2016. Blood DNA yield but not integrity or methylation is impacted after long-term storage. Biopreservation and Biobanking, 14: 1-10.

Chacon-Cortes, D. and Griffiths, L.R. 2012. Comparison of genomic DNA extraction techniques from whole blood samples: A time, cost and quality evaluation study. Molecular Biology Reports, 39: 5961-5966.

Cushwa, W.T. and Medrano, J. F. 1993. Effect of blood storage time and temperature on DNA yield and quality. BioTechniques, 14: 204-207.

Eslami, G., Khalatbari-Limaki S., Ehram-Poush, M.H., Gholamrezaei, M., Hajimohmmadi, B. and Oryan, A. 2016. Comparison of three different DNA extraction methods for Linguatula serrata as a food born pathogen. Iranian Journal of Parasitology, 12: 236-242.

Ghaheri, M., Kahrizi, D., Yari, K., Bbaie, A., Suthar, R.S. and Kazemi, E. 2016. A comparative evaluation of four DNA extraction protocols from whole blood sample. Cellular and Molecular Biology, 62: 119-123.

Gong, R. and Li, S. 2014. Extraction of human genomic DNA from whole blood using a magnetic microsphere method. International Journal of Nanomedicine, 9: 3781-3789. Haque, K.A., Pfeiffer, R.M., Beerman, M.B., Struweing, J.P., Chancock, S.J. and Bergen, A.W. 2003. Performance of high-throughput DNA quantification methods. BMC Biotechnology, 3: 1-10.

Huang, L.H., Li, P.H., Tsai, K.W., Wang, L.J., Huang, Y.H., Kuo, H.C. and Li, S.C. 2017. The effects of storage temperature and duration of blood samples on DNA and RNA qualities. PlosOne, 12: 1-13.

Jain, V., Ratre, Y.K., Amle, D., Mishra, P.K. and Patra, P.K. 2017. Polymorphism of CYP1A1 gene variant rs4646903 and rs1048943 with cervical cancer patients in Chhattisgarh. Environmental Toxicology and Pharmacology, 7: 188-192.

Jelassi, R., Abdallah, R.B., Chelbi, H., Alaya, N.B., Haouet, S., Bouratbine, A. and Aoun, K. 2016. Comparison of two deparaffinization techniques and three DNA extraction methods from paraffin embedded biopsies. Journal of Infection and Molecular Biology, 4: 44-48.

Kim, Y.T., Choi, E.H., Son, B.Y., Seo, E.H., Lee, E.K., Ryu, J.K., Ha., G.W., Kim, J.S., Kwon, M.R., Nam, J.H., Kim, Y.J. and Lee, K.R. 2011. Effects of storage buffer and temperature on the integrity of human DNA. The Korean Journal of Clinical Laboratory Science, 44: 22-30.

Jaya Tiwari et al.

Malentacchi, F., Ciniselli, C.M., Pazzagli, M., Verderio, P., Barraud, L., Hartmann, C.C., Pizzamiglio, S., Weisbuch, S., Wyrich, R. and Gelmini, S. 2015. Influence of pre-analytical procedures on genomic DNA integrity in blood samples: The SPIDIA experience. Clinica Chimica Acta, 440: 205-210.

Qamar, W., Khan, M.R. and Arafah, A. 2017. Optimization of conditions to extract high quality DNA for PCR analysis from whole blood using SDS-proteinase K method. Saudi Journal of Biological Sciences, 24: 1465-1469.

Richardson, A.J., Narendran, N., Guymer, R.H., Vu, H. and Baird, P.N. 2006. Blood storage at 4ºC - Factors involved in DNA yield and quality. Journal of Laboratory and Clinical Medicine, 147: 290-294.

Schuurman, T., Boer, D.R. and Zwet, V.A. 2007. Comparative evaluation of in-house manual, and commercial semi-automated and automated DNA extraction platforms in the sample preparation of human stool specimens for a Salmonella enterica 5′-nuclease assay. Journal of Microbiological Methods, 71: 238-245.

Shams, S.S., Vahed, S.Z., Soltanzad, F., Kafil, V., Barzegari, A., Atashpaz, S. and Barar, J. 2011. Highly effective DNA extraction method from fresh, frozen, dried and clotted blood. BioImpacts, 1: 183-187.

Shimada, T. and Fujii-Kuriyama, Y. 2004. Metabolic activation of polycyclic aromatic hydrocarbons to carcinogens by cytochromes P4501A1 and 1B1. Cancer Science, 5: 1-6. Suguna, S., Nandal, D.H., Kamble, S., Bharatha, A. and Kunkulol, R. 2014. Genomic DNA

isolation from human whole blood samples by non-enzymatic salting out method. International Journal of Pharmacy and Pharmaceutical Sciences, 6: 198-199. Thomas, S.M., Moreno, R.F. and Tilzer, L.L. 1989. DNA extraction with organic solvents in gel barrier tubes. Nucleic Acids Research, 17: 5411.

Wu, D., Li, Y. and Wan, F. 2017. How long can we store blood samples: A systematic review and meta-analysis. EBioMedicine, 24: 277-285.