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Quantitative and Qualitative Chemical Extraction of Deoxyribo Nucleic Acid DNA from Human Cell Organelles

Author Affiliations

  • 1Department of Chemistry, Ravenshaw University, Cuttack - 753003, Odisha, INDIA

Res.J.chem.sci., Volume 4, Issue (8), Pages 75-81, August,18 (2014)

Abstract

The extraction of deoxyribo nucleic acid DNA from biological cell organelles is a complex chemical procedure. The DNA resides in the cell organelles nucleus and mitochondria of biological samples such as blood, bone and tissue. The cell has a cell membrane enclosing the cytoplasm containing cell organelles. The DNA is the self replicating inheriting molecule and encodes all the information of the function of cell organelles. The process of DNA extraction is a chemical cascade of lysing the cell, exposing the DNA to the lysis buffer, separation of DNA from bonded proteins and eluting the DNA in a preservative solution. The cell membrane is lysed in a salt solution of Tris, EDTA and NaCl. The EDTA chelated the inorganic ions of the cell membrane and destabilized it. Tris maintain the pH at 8.0 and also interacted with the lipopolysaccharide of the membrane for further destabilization and hypertonic NaCl solution squeeze out the cell contents. The SDS with polar head and non-polar tail interacted with double layer nuclear and mitochondrial membrane for lysis. The exposed DNA from the cell organelles has boned protein molecules. The serine protease proteinase K digests the protein part of DNA protein complex. The proteins unfolded for interaction of its non-polar groups with non-polar phenol and separated out in the phenol solution. The protein separated DNA solution was mixed with phenol, chloroform and isoamyl alcohol to further denature the residual protein and separate it out with the lipids. The isoamyl alcohol reduces the foaming of phenol and chloroform mix. The DNA was precipitated out from the separated solution by ethanol and sodium acetate. This study investigated the quantity and quality of extracted DNA from cell organelles by spectrophotometric and gel electrophoresis analysis.

References

  1. Kirby K.S., 1957. A new method for the isolation ofdeoxyribonucleic acids: Evidence on the nature of bonds between deoxyribonucleic acid and protein, Biochem. J.66, 495-504 (1957)
  2. Maniatis T., Fritsch E.F., Sambrook J., Molecular Cloning. A Laboratory Manual, (1982)
  3. Miller S.A., Dykes D.D. and Polesky H.F., A simple salting out procedure for extracting DNA from humannucleated cells, Nucleic Acids Res., 16(3)1215 (1988)
  4. Rohland N. and Hofreiter M., Comparison and optimization of ancient DNA extraction, Bio Techniques, 42, 343-352 (2007)
  5. Rohland N. and Hofreiter M., Ancient DNA extractionfrom bones and teeth, Nat. Protocols, 2(7), 1756-1762 2007)
  6. Riemann K., Adamzik M., Frauenrath S., Egensperger R., Schmid K.W., Brockmeyer N.H., Siffert W., Comparison of manual and automated nucleic acid extraction from whole-blood samples, J. Clin. Lab. Anal., 21(4), 244-248 2007)
  7. Lounsbury J.A., Coult N., Miranian D.C., Cronk S.M., Haverstick D.M., Kinnon P., Saul D.J. and Landers J.P., An enzyme-based DNA preparation method for application to forensic biological samples and degraded stains, Forensic Sci. Int. Genet., 6(5), 607-615 (2012)
  8. Sirdah M.M., Super paramagnetic-bead Based Method: An effective DNA extraction from dried blood spots (DBS) for diagnostic PCR, J. Clin. Diagn. Res., 8(4), 1-4 (2014)
  9. Polgár L., The catalytic triad of serine peptidases, Cell. Mol. Life Sci., 62, 2161-2172 (2005)
  10. Negi D.S., Alam M., Bhavani S.A. and Nagaraju J., Multi-step microsatellite mutation in maternally transmitted locus D13S317: A case of maternal allele mismatch in the child, Int. J. of Legal Med., 120, 286-292 (2006)
  11. Shrivastava P., Neetu M., Sharma N.C., Trivedi V.B., Negi D.S. and Verma M.K., Autosomal STR genotyping analysis of juvenile delinquents of Madhya Pradesh: A pilot Study, Adv. Bio. Tech., 13, 20-24 (2013)
  12. Negi D.S., Shrivastava P. and Das S.P., DNA sequencing by polymer synthesis with variable ratio of deoxynucleotide triphosphate and fluorescent dideoxynucleotide triphosphate, Asian J. of Biomed. and Pharma. Sci., 4(32), 32-38 (2014)
  13. Negi D.S. and Das S.P., Computational chemical analysis of DNA sequencing by reducing graphene oxide with the released H ion during polymer synthesis, J. of Chem. and Pharma. Res., 6(7), 2190-2196 (2014)
  14. Sahoo S. and Kashyap V.K., Genetic variation at 15 autosomal microsatellite loci in three highly endogamous tribal populations of Orissa, India, Forensic Sci. Int., 130, 189-193 (2002)
  15. Sahoo S. and Kashyap V.K., Influence of language and ancestry on genetic structure of contiguous populations: A microsatellite based study on populations of Orissa, BMC Genet., 6(4), 2156-2165 (2005)
  16. Almahy H.A., Ali M.A. and Ali A.A., Extraction of carotenoids as natural dyes from the daucus carota linn (carrot) using ultrasound in kingdom of saudi arabia, Res. J. Chem. Sci.,3(1), 63-66 (2013)
  17. Shukla H.M., Solanki Y.K., Shah A.R., Shah P.J., Shah A.I. and Raj D.S., Synthesis, characterization and antimicrobial studies of coordination polymers, Res. J. Chem. Sci.,3(1), 48-56 (2013)
  18. Jadhav U.M., Patel S.N. and Patil R.S., Synthesis of silver sulphide nanoparticles by modified chemical route for solar cell applications, Res. J. Chem. Sci.,3(7), 69-74 2013)
  19. Manjula B., Arul A.S., Antony S.A., Studies on DNA cleavage and antimicrobial screening of transition metal complexes of 4-aminoantipyrine schiff base, Res. J. Chem. Sci.,3(12), 22-28 (2013)
  20. Pravin S.B., Sakharam B.D. and Umakant B.C., Synthesis and antimicrobial screening of chalcones containing imidazo [1,2-a] pyridine nucleus, Res. J. Chem. Sci.,3(12), 38-42 (2013)