Isolation and characterization of high quality DNA from marine benthic macroalgae.
The isolation of high quality DNA is essential for many molecular biology applications including polymerase chain reaction (PCR) and endonuclease restriction digestion based techniques. An easy and inexpensive protocol has been developed for extracting genomic DNA from seven species of algae viz. Lola capillaries, Enteromorpha intestinalis, Ulva lactuca and Rhizoclonium sp belonging to Chlorophyceae, Catenella nipae, Polysiphonia mollis belonging to Rhodophyceae and Dictyota ceylanica belonging to Phaeophyceae group were collected from the coastal regions of Sunderban delta in West Bengal, India dominantly growing on mud flats, bark of different mangrove trees, pneumatophores, stilt roots, concrete surfaces, wooden and bamboo poles, sides of the boats and other water vehicles inundated during high tides. The DNA was found suitable for restriction endonuclease digestion and PCR amplification with randomely amplified polymorphic DNA (RAPD) primers. The A260/A280 ratio of 1.15 0.14 to 1.94 indicated little contamination from proteins and polysaccharides. The PCR amplification with RAPD primers showed its suitability in PCR based techniques and the restriction digestion with Eco RV confirmed its suitability for hybridization based techniques. The protocol is equally good for isolating DNA from both fresh as well as preserved materials.
Chakraborty
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N.V. Patel College of Pure and Applied Sciences, Vallabh Vidyanagar 388 120, India. su_kalyanc@yahoo.co.uk
Vijayan
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Nair
C V
CV
Santra
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SC
Bhattacharya
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T
D048028:DNA, Algal / Q000302:isolation & purification* / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific; D056890:Eukaryota / Q000235:genetics*; D023281:Genomics / Q000379:methods*; D016133:Polymerase Chain Reaction
D048028:DNA, Algal; D015252:Deoxyribonucleases, Type II Site-Specific; C027440:GATATC-specific type II deoxyribonucleases