Modifying and adapting a plant-based DNA extraction protocol for human genomic DNA extraction: a cost effective approach

M.E Kooffreh,O.U Udensi,A.J Umoeyen

doi:10.4314/gjpas.v23i1.1

Abstract

This research involved modification of a standard protocol for DNA extraction in plants for its suitability as an extraction method for DNA in human whole blood in which some of the plasma was removed. Human blood samples were obtained from a 100 apparently healthy individuals residing in Calabar. The modified DNA procedure yielded good quality genomic DNA which was used in carrying out allele specific polymerase chain reaction which also yielded good quality amplicons. This method is simple and suitable for the extraction of DNA from human red cell.

Highlights

The extraction of human genomic DNA is obviously a crucial step in molecular techniques such as genotyping, DNA fingerprinting, diagnostics and forensic analyses
Maurya et al (2013) comparing salt-out as modified, phenol-chloroform and DNA extraction kit method showed that modified salt-out protocol gave higher human genomic DNA yield of 40.8μg/ml followed by phenol-chloroform methods (38.5 μg/ml) and kit method (35.3μg/ml)
Our result showed that the modification of the Dellaporta et al.’s protocol for use in the genomic DNA extraction from packed human red blood cells yielded good quality DNA

Summary

Introduction

The extraction of human genomic DNA is obviously a crucial step in molecular techniques such as genotyping, DNA fingerprinting, diagnostics and forensic analyses. The implication of the above is that a cost effective, safe, rapid and reliable DNA extraction protocol becomes imperative for use, especially in smaller laboratories. Many economic and in-house methods for DNA extraction from mammalian peripheral blood exist namely salt-out technique (Miller et al, 1988), phenol-chloroform extraction (Sambrook et al, 1989) and QIAamp DNA Blood Kits methods (Maurya et al, 2013). Maurya et al (2013) comparing salt-out as modified, phenol-chloroform and DNA extraction kit method showed that modified salt-out protocol gave higher human genomic DNA yield of 40.8μg/ml followed by phenol-chloroform methods (38.5 μg/ml) and kit method (35.3μg/ml)

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Conclusion