Adebayo Liasu Ogunkanmi
1
*, Bola Oboh
1
, Bukola Onifade
2
, Adeniyi Adewale
Ogunjobi
3
, Idowu Adewumi Taiwo
1
, Oluwatoyin Temitope Ogundipe
2
1
Cell Biology and Genetics Department, University of Lagos, Lagos, Nigeria,
2
Botany and Microbiology Department, University of Lagos, Lagos, Nigeria
3
Botany and Microbiology Department University of Ibadan, Ibadan, Nigeria
*Corresponding Author: bayogun2000@yahoo.com
An improved method of extracting genomic DNA
from preserved tissues of Capsicum annuum for
PCR amplification
Abstract
In this study we present a method for the extraction of genomic DNA from the different tissues
of the Pepper (Capsicum annuum). A standard protocol of Dellaporta was reviewed and
modified for DNA extraction from the preserved tissues of the Capsicum sample which was
believed to contain high level of polysaccharides. The modified protocol employed yielded a
high quality DNA and was found to be suitable for PCR and RAPD analyses. The procedure
was also found to be reliable and suitable where some materials are not available and does not
require phenol-chloroform extraction. The method also allowed for the preservation of plant
tissues for some days from a locality where storage facilities are not accessible. We also
discovered that irrespective of the sources of tissues, a good quality DNA was obtained. The
quantity of DNA produced from the fleshy mesocarp tissue was more than the quantity
obtained from the seeds of the same weight, this is probably due to the hard nature of the
seeds and there may have been no complete breakdown of the cell wall to release the cellular
contents.
Keywords: Capsicum, genomic, mesocarp, pepper, polysaccharides.
Ogunkanmi AL, Oboh B, Onifade B, Ogunjobi AA, Taiwo IA, Ogundipe OT (2008) An improved
method of extracting genomic DNA from preserved tissues of Capsicum annuum for PCR
amplification. EurAsia J BioSci 2, 14, 115-119.
www.ejobios.com/content/2/14/115-119
©EurAsian Journal of BioSciences, 2008 115
EurAsian Journal of BioSciences
EurAsia J BioSci 2, 115-119 (2008)
Molecular techniques require isolation of
genomic DNA of suitable purity. The growing
number of DNA extraction protocols for
specific plant species are not always simple
and cannot be reproduced for all species
(Porebski et al. 1997).
Polysaccharide contamination is a common
problem in higher plant DNA extraction. DNA
samples are often contaminated with Melicera
colloidal Lyalosome, which cannot be
dissolved in water or TE buffer (Yun-Jiang et
al. 2003).
Plant contaminants like polysaccharides
and phenolic compounds are difficult to
separate from DNA and are readily identified
as they impart a sticky gelatinous brown color
to the DNA isolated and interfere with
polymerases, ligases and restriction enzymes
(Fang et al. 1992, Michard et al. 1995,
Porebski et al. 1997, Csaikl et al. 1998,
Tribouch et al. 1998, Belletti et al. 1998,
Schlink and Reski, 2002). These
contaminants are in abundance in the foliage
of perennials, and they co-extract with the
DNA (Scott and Playford 1996, Shephered et
al. 2002, Bhattarcharjee et al. 2004).
DNA extraction becomes more difficult
when working with perennials or tree plant
species. Several factors are known to limit the
isolation of pure DNA from such species that
are rich in impurities; such as terpernes,
polyphenols, and polysaccharides. Such
factors include the amount of tissue available,
the condition of the plant material, the
numbers of steps involved in the extraction
procedure and the required purity level (Helen
Received : August, 2008
Accepted : December, 2008
Printed : December, 2008
INTRODUCTION
Research Note