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KMID : 0613819990090010015
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Journal of Life Science
1999 Volume.9 No. 1 p.15 ~ p.21
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Limits of Direct PCR Amplication from Seaweeds Using Arvitrary and ITS Primers
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Abstract
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The random amplified polymorphic DNAs (RAPD) assay is a simple and useful tool in identification of apppropriate genetic markers, that requires no knowledge of target DNA sequence. RAPD products were generated directly from seaweed tissues, without prior nucleic acid extraction, of Porphyra yezoensis, Ulva pertusa and Undaria pinnatifida. The nuclear rDNA internal transcribed spacer (ITS) fragment however was not amplified directly from the seaweed tissues. Using DNA extracted by the LiCl method, both the ITS and RAPD´s have been amplified by the polymerase chain reaction. RAPD of P. yezoensis, thallus (n) and conchocelis (2n) produced lots of different polymorphic bands (36-50%) depending on the arbitrary primers used. Difference was also observed between direct tissues amplification and DNA extracts amplification (53-57%). Thus it is important to use the same ploidy of tissue for DNA extraction and as a RAPD template.
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