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An optimized mini-preparation method
to
obtain high-quality genomic DNA from
mature leaves of sunflower
J.T. Li1, J. Yang1,2, D.C. Chen1,
X.L. Zhang1 and Z.S. Tang1
1School of Life Sciences, China
West Normal University, Nanchong,
Sichuan, P.R. China
2Institute of Rare Animals and Plants, China West
Normal University,
Nanchong, Sichuan, P.R. China
Corresponding author: J. Yang
E-mail: yangjunlz@tom.com
Genet. Mol. Res. 6 (4): 1064-1071 (2007)
Received June 18, 2007
Accepted October 24, 2007
Published December 4, 2007
ABSTRACT. In order to investigate the
mutation characteristics and to further examine the genetic
variation of mutant sunflower (Helianthus annuus) obtained
in plants grown from seeds exposed to space conditions, only
the mature tissues such as leaf and flower could be used for
DNA extraction after mutation characteristics were confirmed.
The rich contents of polysaccharides, tannins, secondary metabolites,
and polyphenolics made it difficult to isolate high-quality
DNA from mature leaves of sunflower according to previous reports.
Based on the comparison of the differences in previously reported
protocols, a modified method for the extraction of high-quality
DNA was developed. Using this protocol, the DNA isolated from
sunflower was high in quality and suitable for restriction digestion
(EcoRI, HindII, BamHI), random amplified
polymorphic DNA study and further molecular research. Therefore,
the modified protocol was suitable for investigating the genetic
variation of sunflower using mature leaf DNA.
Key words: CTAB, DNA extraction protocol,
PVP, RAPD, Sunflower
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