title: An efficient method for large-scale isolation of plasmid DNAs by heat-alkali co-denaturation.
abstract: A method is presented for efficient and large-scale isolation of plasmid DNAs from bacterial cells. Based on the cooperativity of heat and alkali actions, the method provides DNA preparations with high quality and yield (about 2 micrograms of DNA/ml culture), which are completely digestable by restriction enzymes and have a high transformation efficiency. Furthermore, the DNA preparations are extremely stable, and even through 4-year storage at -20 degrees C, the electrophorogram and transformation efficiency remain as high as before. The factors affecting the stability of various DNA samples are discussed.
authors.affiliation: Department of Biochemistry, Nanjing University, People's Republic of China.
authors.forename: N E
authors.initials: NE
authors.lastname: Sun
authors.forename: B H
authors.initials: BH
authors.lastname: Shen
authors.forename: J M
authors.initials: JM
authors.lastname: Zhou
authors.forename: J
authors.initials: J
authors.lastname: Yuan
authors.forename: X X
authors.initials: XX
authors.lastname: Xu
authors.forename: D X
authors.initials: DX
authors.lastname: Zhu
authors.forename: K K
authors.initials: KK
authors.lastname: Han
mesh_terms: D000433:1-Propanol; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004492:Edetic Acid; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D010957:Plasmids / Q000302:isolation & purification*
chemical_list: D004269:DNA, Bacterial; D000433:1-Propanol; D004492:Edetic Acid