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Extraction of plasmid-like DNA and high-quality total DNA from Porphyra yezoensis
Authors:Guo Baotai  Dai Jixun  Shen Songdong  Bi Yuping  Shan Lei and Li Guangcun
Institution:1.College of Marine Life Sciences, Ocean University of Qingdao, Qingdao 266003, China2.Biotechnology Research Center, Shandong Academy of Agricultural Sciences, Jinan 250100
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.
Keywords:Porphyra yezoensis  somatic cells  total DNA  glassmilk  restriction digestion  plasmid like DNA
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