ATP™ Plasmid Mini Kit is designed for the rapid , easily-handling, and cost-effective isolation of plasmid or cosmid DNA from 1~4 ml of bacterial cultures. For large scale plasmid purification we recommend ion exchange based Fast Ion Plasmid Mini/Maxi Kits.
1. This procedure uses modified alkaline lysis methodand RNase treatment for creating cleared cell lysate with minimal genomic DNA and RNA contaminants.
Subsequently, the lysate is neutralized and adjusted to high-salt binding conditions adaptable to the adsorption of DNA in one step. In the presence of a chaotropic salt, the plasmid DNA in the lysate binds to the uniquely-designed glass fiber matrix in the spin column .
Whereas RNA, cellular proteins, and other unwanted impurities flow through the column and are easily and efficiently removed from reaction mixture . After a brief washing step with ethanol-contained wash buffer to remove endonucleases, salts and other contaminants .
4. The purified plasmid DNA is eluted by a low salt elution buffer or water. The entire procedure can be completed in 20 ninutes and the purified plasmid DNA is immediately ready for restriction digestion, ligation, PCR, and sequencing.
The procedure does not require DNA phenol extraction and alcohol precipitation.