Principle

The classic anion exchanger for purification of transfection-grade plasmid DNA!

Bacteria harvested from the culture broth are lysed by using a modified alkaline/SDS procedure. Both chromosomal and plasmid DNA are denatured under these alkaline conditions. Potassium acetate is then added to the denatured lysate, which causes the formation of a precipitate containing chromosomal DNA and other cellular compounds and neutralizes the lysate. Plasmid DNA, which remains in solution, can revert to its native supercoiled structure. After equilibrating the appropriate NucleoBond® AX column the cleared lysate is loaded onto the column and plasmid DNA is bound to the anion-exchange resin. After subsequent washing steps, the purified plasmid DNA is eluted in a high-salt buffer and precipitated with isopropanol. Finally, the plasmid DNA is reconstituted in TE buffer for further use.

For convenience of the customer, NucleoBond® PC kits contain NucleoBond® AX columns, all required buffers, reagents, and accessories. The NucleoBond® plasmid purification kits are available in several sizes to accommodate a wide range of purification needs.

Features

* High-copy plasmids
** Low-copy plasmids