An improved method has been developed for the large-scale purification of covalently closed circular (CCC) plasmid DNA molecules of sizes ranging from 4.3 to 73 kb. This protocol uses an alkaline-lysis procedure followed by acid-phenol extraction but with several modifications to previously reported methods. The principal modification is the replacement of NaCl by MgCl2 in the extraction buffer to improve yield and to remove chromosomal and other non-CCC plasmid DNA. Plasmid DNA can be purified in less than 1 h and used successfully in restriction enzyme analysis and cloning experiments.
机构:
Peking Univ, Sch Life Sci, Grad Program, Beijing, Peoples R China
Natl Inst Biol Sci, 7 Sci Pk Rd,ZGC Life Sci Pk, Beijing, Peoples R ChinaPeking Univ, Sch Life Sci, Grad Program, Beijing, Peoples R China
Gao, Zhenchao
Yan, Liwei
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Natl Inst Biol Sci, 7 Sci Pk Rd,ZGC Life Sci Pk, Beijing, Peoples R ChinaPeking Univ, Sch Life Sci, Grad Program, Beijing, Peoples R China
Yan, Liwei
Li, Wenhui
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Natl Inst Biol Sci, 7 Sci Pk Rd,ZGC Life Sci Pk, Beijing, Peoples R China
Tsinghua Univ, Tsinghua Inst Multidisciplinary Biomed Res, Beijing, Peoples R ChinaPeking Univ, Sch Life Sci, Grad Program, Beijing, Peoples R China