Abstract
A chromatographic method for the rapid isolation of preparative amounts of plasmid DNA without the use of cesium chloride centrifugation is described. The protocol uses the alkaline extraction procedure and an exclusion column of Fractogel TSK 75S. From a clear lysate it is possible to obtain plasmid DNA completely free of proteins, RNA, and chromosomal DNA. From partially purified plasmid the procedure allows the separation of the different forms. This technique was successfully applied to different plasmids ranging in size from 2.9 to 17.5 MDa. It is a preparative method yielding easily 500 μg of pBR322 from 1 liter of amplified culture. The plasmid is suitable for topoisomerase I, topoisomerase II, and EcoRI assays.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
