The E.Z.N.A.^® Fastfilter Plasmid Maxiprep Kit offers purification of high quality plasmid DNA in less than 30 minutes. The E.Z.N.A.^® Fastfilter Plasmid Maxiprep Kit features a filter syringe for lysate clearance and HiBind^® DNA Maxi-columns that can be used with vacuum or spin purification formats.

Principle

The E.Z.N.A.^® Fastfilter Plasmid Maxiprep Kit combines the power of HiBind™ technology with the time-tested consistency of alkaline-SDS lysis of bacterial cells to deliver high quality DNA. HiBind™ matrix that avidly, but reversibly, binds DNA under certain optimal conditions allowing proteins and other contaminants to be removed. Nucleic acids are easily eluted with de-ionized water or low salt buffer. This kit also include a special filter cartridge, which replaces the centrifugation step following alkaline lysis. Following lysis the DNA is bound to the silica membrane and contaminants are removed with a simple wash step. The use of maxi-columns facilitate the binding, washing, and elution steps thus enabling multiple samples to be simultaneously processed. For more information on columns, click Flexible Multi-Format Columns.

Procedure

Pellet cells from ~ 200 ml of overnight culture. Re-suspend collected cells in Solution I / RNase I. Add Solution II and gently mix by inverting and rotating the tube to obtain a cleared lysate. Add neutralization buffer and gently mix by inverting several times until a flocculent white precipitate forms. Add GBT Buffer and gently mix by inverting 1-2 times. Immediately pour the lysate into the barrel of the Lysate Clarence Filter Syringe. Allow the cell lysate to sit and pass through the filter. Collect the cleared cell lysate in a new tube. Equilibrate the HiBind^® DNA maxi column with GPS buffer and add the cleared lysate to it. Centrifuge at 3000 x g for 2 minutes. Discard the flow-through and wash the column once with HB buffer and twice with wash buffer. Elute plasmid DNA using sterile de-ionized water or TE buffer for further use.

Applications

E.Z.N.A.^® Fastfilter Plasmid Maxiprep Kit provides high quality, reproducible plasmid DNA for use in the following applications :

• Automated fluorescent DNA sequencing
• Transfection of Mammalian cells
• Restriction Digestion