Macherey-Nagel™ Buffer N2

Description

NucleoBond™ Plasmid purification kits incorporate NucleoBond™ AX technology, a silica-based anion exchanger developed for routine separation of different classes of nucleic acids, allowing superior biological activity of the purified nucleic acids and rendering high mass recoveries possible by minimising non-specific adsorption effects. Bacteria harvested from the culture broth are lysed 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, causing the formation of a precipitate containing chromosomal DNA and other cellular compounds, thus neutralising 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 applied 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. Alternatively, NucleoBond™ Finalizer can be used for concentration and desalination of eluted plasmid DNA. NucleoBond™ folded filters (included in the Midi (PC 100), Maxi (PC 500), Mega (PC 2, 000) and Giga (PC 10,000) kits) eliminate the centrifugation step after the alkaline lysis. The filters reduce “hands-on” time and eliminate plastic waste common to other filtration systems. In approximately 2min (Midi) and 10min (Maxi) of unattended operation, complete removal of SDS and cellular debris from plasmid samples is achieved. NucleoBond™ folded filters do not induce shearing of large DNA constructs, such as PACs or BACs. Kit types and their components: NucleoBond™ Mini (PC 20) - NucleoBond™ AX 20 columns, buffers, Rnase A. NucleoBond™ Midi (PC 100) - NucleoBond™ AX 100 columns, buffers, Rnase A folded filters. NucleoBond™ Maxi (PC 500) - NucleoBond™ AX 500 columns, buffers, Rnase A, folded filters XL. NucleoBond™ Mega (PC 2000) - NucleoBond™ AX 2000 columns, buffers, Rnase A, folded filters XL. NucleoBond™ Giga (PC 10,000) - NucleoBond™ AX 10,000 columns, buffers, Rnase A, folded filters. For further information on this product contact Customer Services.

• NucleoBond™ folded filters eliminate the centrifugation step for easy, fast and convenient clarification of bacterial lysate
• Columns run by gravity flow, allowing purification of multiple samples in parallel
• Working range from nanograms to milligrams DNA
• Recovery of plasmid DNA is >90%
• Transfection-grade DNA
• High batch-to-batch reproducibility
• No use of organic solvents
• Alcohol precipitation of DNA/RNA High and low copy number plasmid DNA purifaction fromE. colithat is suitable for transfection, sequencing and cloning.