Open AccessA new method for the purification and identification of covalently closed circular DNA molecules
Author(s) -
Michael Zasloff,
Gordon D. Ginder,
Gary Felsenfeld
Publication year - 1978
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/5.4.1139
Subject(s) - biology , dna , circular dna , identification (biology) , computational biology , covalent bond , base sequence , genetics , gene , genome , botany , physics , quantum mechanics
A new technique has been developed for the rapid isolation of covalently closed circular DNA molecules. The procedure is a selective extraction based on differences in the partitioning of covalently closed circular DNA molecules and noncovalently closed species between phenol and water at acid pH and low ionic strength. Under the conditions described, linear as well as nicked circular DNA is extracted into phenol, while covalently closed circular DNA molecules remain in the water phase. The method permits the quantitative isolation of covalently closed circular DNA from either total cellular DNA or partially purified preparations, to a degree of purity comparable with buoyant density procedures.