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ALGAE > Volume 21(2); 2006 > Article
ALGAE 2006;21(2): 261-266. doi: https://doi.org/10.4490/algae.2006.21.2.261
A Simple Method for Extraction of High Molecular Weight DNA from Porphyra Tenera (Rhodophyta) Using Diatomaceous Earth
Tae Hoon Kim1, Mi Sook Hwang2, Ju-Dong Song1, Min-Hyuk Oh1, Yong-Hwan Moon1, Ik Kyo Chung3, Tae-Hyoung Rhew4 and Choon-Hwan Lee1*

1Department of Molecular Biology, Busan National University, Busan 609-735, Korea
2Seaweed Research Center, South Sea Fisheries Research Institute, National Fisheries Research and Development Institute, Mokpo 530-831, Korea
3Department of Marine Science, Busan National University, Busan 609-735, Korea
4Department of Biology, Busan National University, Busan 609-735, Korea
ABSTRACT
The innate soluble polysaccharides and phenolic compounds of marine macroalgae are serious contaminants which interfere with experimental procedures such as restriction enzyme digestion, polymerase chain reaction (PCR) and other enzymatic reactions using extracted DNA samples. The viscous polysaccharides are co-precipitated with DNA samples by isopropanol or ethanol precipitation in conventional experiment. To overcome the problem, a method for the isolation of high molecular weight DNA from Porphyra tenera is developed with the application of diatomaceous earth column. The isolated DNAs by this method were about 50-100 kb in size and could be digested well with restriction enzymes. The nuclease activity seemed to be minimal, and high reproducibility in the arbitrary primed PCR for RAPD analyses was a distinctive feature. These results suggest that this method is very efficient in isolating nucleic acid from macroalgae including Porphyra.
Key words: Diatomaceous earth, DNA preparation, Porphyra tenera, RAPD pattern, Rhodophyta


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