Detergents in molecular biology: DNA extraction and purification Chapter uri icon

abstract

  • Over the last 20 years, epidemiologist have shifted their attention from the analysis of phenotypic features to evaluating the way genetic traits determine disease causation and severity. This trend has clearly benefitted from the coming of age of versatile molecular based techniques such as the Polymerase Chain Reaction (PCR) and the now widespread availability of high-throughput molecular screening assays. In this context, deoxyribonucleic acid (DNA) has rapidly become the biospecimen of choice as it is readily obtained, transported, processed and stored. Both molecular epidemiology and genomic characterization studies require the typing of large numbers of specimens in order to achieve statistical significance and enormous advances, such as microarray technology, have resulted as a consequence of this. Nevertheless a usually overlooked yet crucial aspect of nucleic acid based typing is the DNA extraction process. DNA extraction is the process by which nucleic acids are retrieved from their biological compartments and deprived of proteins, salts and other substances that might hinder their analysis. In their natural state nucleic acids are compartmentalized inside the cell nucleus or mitochondrion and protected by proteins and a lipid envelope. In order to extract these nucleic acids cells must be burst open and the contaminating proteins and lipids eliminated. Detergents of essentially every type have been used to disaggregate the lipid components of cell membranes and are common components of essentially all DNA extraction methods. In this chapter a brief historical review of the different DNA extraction techniques that have been developed is made followed by a detailed presentation of the most relevant applications of detergent-based DNA extraction techniques. Finally, we describe our experience at implementing and optimizing a simple and low-cost laundry-detergent-based DNA extraction method for the extraction of high-quality human genomic DNA from peripheral blood samples and the many uses that such material has been put to use for. © 2010 Nova Science Publishers, Inc.

publication date

  • 2011-01-01