Description
The E.Z.N.A.® FastFilter Plasmid DNA Mini Kit is designed for rapid purification of high-quality plasmid DNA from up to 5 mL of culture following the alkaline-lysis method. The kit features an innovative and first-of-its-kind FastFilter Mini Column nested within a regular HiBind® DNA Mini Column to combine lysate clearance and DNA binding into one or two centrifugation steps depending on the culture volume. The protocol time is just 9 minutes when starting with bacterial pellet from up to 3 mL culture volume. The protocol time for culture volumes greater than 3 mL will be slightly longer due to two centrifugation steps for lysate clearance. Following the wash and elute steps, purified plasmid DNA is immediately ready for various downstream applications such as routine screening, restriction enzyme digestion, transformation, PCR, and DNA sequencing.
Typically, up to 5 mL overnight bacterial culture in LB or other suitable growth medium is centrifuged to pellet the bacterial cells. The cells are then lysed under alkaline conditions and subsequently neutralized to form white flocculent precipitate. Neutralized lysate is then transferred to a FastFilter Mini Column nested within a HiBind® DNA Mini Column. Upon adjustment of binding conditions, the lysate is centrifuged through this FastFilter HiBind® configuration to both clear the lysate and facilitate binding of DNA to the HiBind®
DNA Mini Column. The FastFilter Mini Column is discarded, and the HiBind® DNA Mini Column is subjected to two wash steps to remove salt and other contaminants. High-quality plasmid DNA is then eluted in low-salt elution buffer and is ready for use in a wide range of downstream applications mentioned above.
