A Simple Method for Extraction of High Molecular Weight DNA from Porphyra Tenera (Rhodophyta) Using Diatomaceous Earth

doi:2006.21.2.261

Algae > Volume 21(2); 2006 > Article

Algae 2006;21(2): 261-266. doi: https://doi.org/10.4490/algae.2006.21.2.261

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