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Indian Journal of Biotechnology (IJBT) >
IJBT Vol.11 [2012] >
IJBT Vol.11(3) [July 2012] >
| Title: | A simplified high yielding miniprep genomic DNA extraction protocol for three chemotypically different plant species |
| Authors: | Adhikari, S
Chattopadhyay, S K
Ghosh, P D |
| Keywords: | DNA extraction
PCR
Phenol-chloroform
Restriction digestion
SDS |
| Issue Date: | Jul-2012 |
| Publisher: | NISCAIR-CSIR, India |
| Abstract: | The presence of metabolites has been observed to create interference
with DNA isolation procedures and downstream reactions, such as, DNA
amplification, restriction and cloning. Moreover, the chemotypic heterogeneity
among species may cause hindrance in optimal DNA yields with a single protocol.
Thus, even closely related species may require different isolation protocols. A
rapid and high yielding DNA extraction procedure from heterogeneous plant tissues of 3 chemotypic different plants has
been presented here. This modified
method required grinding of plant tissues in DNA extraction buffer [200
mM Tris-HCL
(pH 8.0), 200 mM NaCl and 25 mM EDTA,
and 1% PVP], followed by phenol:chloroform:isoamyl alcohol (25:24:1, v/v/v)
treatment. Ethanol in the presence of high salt concentration was then used to
precipitate the DNA and for removal of polysaccharides. The average yield of
DNA ranged from 1.68 to 5.37 mg g-1 of plant tissue and the purity
was 1.6 to 1.9. The DNA was quite suitable for PCR using RAPD and also for
restriction digestion. This method did not require liquid nitrogen for
fixation, RNase treatment or storage at –80°C, making it advantageous over other common protocols.
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| Page(s): | 337-340 |
| CC License: |  CC Attribution-Noncommercial-No Derivative Works 2.5 India |
| ISSN: | 0975-0967 (Online); 0972-5849 (Print) |
| Source: | IJBT Vol.11(3) [July 2012]
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