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

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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.

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