This protocol describes an easy, convenient and rapid method of genomic DNA isolation and purification from Gram (+) and (-) bacterial broth suspensions. The workflow, dubbed as a 1-step, single buffer system is a reversal of the traditional silica membrane and magnetic bead-based DNA extraction protocols. Proteins, cellular debris and other low-molecular weight contaminants are retained by the nexttec\u2122 sorbent while pure dsDNA is pulled through as an eluate in a single wash step.The protocol described is suitable for extraction of DNA from pure bacterial isolates of the following organisms:1. Acinetobacter baumannii2. Escherichia coli3. Klebsiella pneumoniae4. Staphylococcus aureus5. Neisseria gonorrhoea6. Salmonella typhi and typhimurium7. Non-typhoidal Salmonella spp.8. Pseudomonas aeruginosa9. Streptococcus pneumoniae10. Vibrio choleraeThis protocol has been adapted from nucleic acid extraction methods developed by nexttec Biotechnologie GmbH.This is an open-access protocol distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike.