Extraction of plasmid-like DNA and high-quality total DNA from Porphyra yezoensis |
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Authors: | Guo Baotai Dai Jixun Shen Songdong Bi Yuping Shan Lei Li Guangcun |
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Affiliation: | 1.College of Marine Life Sciences, Ocean University of Qingdao, Qingdao 266003, China2.Biotechnology Research Center, Shandong Academy of Agricultural Sciences, Jinan 250100 |
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Abstract: | Somatic cells were prepared from sea snail enzyme digests of Porphyra yezoensis thalli. Us ing SDS - Proteinase K as extraction solution, total DNA was isolated from the somatic cells. The crude extracts of total DNA were purified with glassmilk, and the resulting DNA was of sufficient quality for digestion of restriction endonuclease. DNA bands were clearly observed in the restriction patterns of EcoRI, PstI and HaeIII respectively. The presence of DNA hands in the restriction pattern of total DNA indicated that the genome of Porphyra yezoensis may be small. Unexpectedly, using guanidinium isoth iocyanate and sarcosyl as extraction solution, a plasmid-like DNA band (2.3 Kb) was directly found in the isolated total DNA of Porphyra yezoensis. A very simple and convenient method for plasmid-like DNA isolation has been established. |
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Keywords: | Porphyra yezoensis somatic cells total DNA glassmilk restriction digestion plasmid like DNA |
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