Isolation of bacterial artificial chromosome DNA by means of improved alkaline lysis and double potassium acetate precipitation

Abstract

This work describes an easy alkaline lysis method for isolatating bacterial artificial chromosome (BAC) DNA in sufficient quantity and quality for further manipulation without the need to use a kit. The method starts with 10 mL of culture and, by alkaline lysis only, renders up to 150 ng of DNA per milliliter of culture. This BAC DNA was successfully digested with restriction enzymes, sequenced, and subjected to PCR.