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Research Journals >
Indian Journal of Biotechnology (IJBT) >
IJBT Vol.09 [2010] >
IJBT Vol.09(2) [April 2010] >
| Title: | An efficient genomic DNA isolation protocol for RAPD and SSR analysis in Acorus calamus L. |
| Authors: | Ginwal, H S
Mittal, Neha |
| Keywords: | Acorus calamus CTAB
DNA extraction
PVP
RAPD
SSR |
| Issue Date: | Apr-2010 |
| Publisher: | CSIR |
| Abstract: | Molecular based genetic analysis requires good quality of DNA in sufficient
quantity to generate robust DNA fingerprints. The leaves of sweet flag (Acorus calamus L.) contain high amounts
of polyphenolic compounds and polysaccharides, which interfere in the
amplification reactions. Four genomic DNA extraction methods were tested for
yield, quality and suitability of genomic DNA for RAPD (random amplified
polymorphic DNA) and SSR (simple sequence repeat) marker analysis in sweet
flag. Fresh young leaves were subjected to the already available protocols and
a procedure was devised after modification in the protocol of Stange et al for the isolation of total genomic
DNA. The modified method employs high concentrations of polyvinyl pyrrolidone,
addition of lithium chloride solution as well as additional washing step of DNA
pellets. The yield was found approximately 200-500 µg
DNA per 100 mg of plant material and the purity ratio was found 1.7-2.0.
Usage of highly concentrated PVP extraction buffer and
chloroform:isoamylalcohol repetition step was found useful to overcome problems
from polyphenolic compounds and polysaccharides. The extracted DNA through this
method was found suitable for RAPD and SSR analysis. |
| Page(s): | 213-216 |
| ISSN: | 0975-0967 (Online); 0972-5849 (Print) |
| Source: | IJBT Vol.09(2) [April 2010]
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