The levels of yield and purity of genomic DNA from five tomato cultivars subjected to two DNA extraction techniques

S A Ganiyu,J O Agbolade,J E Imonmion,S M Yusuf

doi:10.4314/njb.v33i1.19

Abstract

Isolation of good quality genomic DNA from different plant materials is an important prerequisite for many molecular techniques related to both basic and applied research in the areas of plant molecular biology, crop improvement, biodiversity studies and conservation of genetic materials. Therefore, the need to extract reasonably pure DNA of both good quality and quantity for the downstream successful DNA-based diagnostic techniques is required. In this study, yield and purity of the extracted DNAs, using Dellaporta et al. (1983) and the use of Zymo Research Plant/Seed DNA MiniPrep Kit (ZR kit), were quantified with Nanodrop spectrophotometer and on agarose gel after polymerase chain reaction (PCR) amplification. The results of the DNA yield and purity were in the range of 24.42 to 1430.52 ng/μl and 1.11 to 2.00, respectively. The outstanding DNA yield in Kerewa cultivar in yield (1430.52 ng/μl) and purity (1.99) makes it appropriate for downstream DNA-based diagnostic techniques in biotechnology research. On agarose gel DNA bands appeared to be compact with negligible smearing with Dellaporta et al. method indicating good quality of DNA. The results demonstrated that good yield of very pure, intact, high quality genomic DNA could be obtained from tomato leaves using the method of Dellaporta et al. Keywords : DNA, extraction, purity, tomato, yield.

Highlights

DNA extraction is a routine step in many biological studies including molecular identification, phylogenetic inference, genetics, and genomics
It was observed that Kerewa had a unique DNA yield (1430.52 ng/μl) above other cultivars using Dellaporta et al protocol; with Purewater cultivar produced the least yield of 687.51 ng/μl
DNA can interfere polymerase chain reaction (PCR) reactions, for example polyphenolic compounds, polysaccharides and RNA (Krishna et al, 2012) and these can hamper the isolation of good quality DNA (Arif et al, 2010)

Summary

Introduction

DNA extraction is a routine step in many biological studies including molecular identification, phylogenetic inference, genetics, and genomics. Ecology of southwestern Nigeria is very favorable for tomato production It is an economic fruit vegetable for tomato growers, consumed fresh and used to manufacture a wide range of processed products. One of the most challenging is bacterial wilt caused by Ralstonia solanacearum. This disease can constitute up to 100% yield loss in endemic areas (Popoola et al, 2015). Breeding a resistant variety using an updated gamplasm as a donor typically requires a series of backcrosses to the cultivated recurrent parent to combing desirable characteristics. The application of molecular breeding through extraction

Methods

Results

Conclusion