The High-Speed Plasmid Mini Kit was designed for both plasmid and cosmid DNA purification. This plasmid kit will purify plasmid DNA/cosmid DNA from 1-5 ml of cultured bacterial cells using an efficient plasmid miniprep system. The High-Speed Plasmid Mini Kit uses a modified alkaline lysis method and RNase treatment to obtain clear cell lysate with minimal genomic DNA/RNA contaminants. In the presence of chaotropic salt, plasmid DNA in the lysate binds to the glass fiber matrix of the plasmid spin column. Contaminants are removed with a Wash Buffer (containing ethanol) and the purified plasmid DNA is eluted by a low salt Elution Buffer, TE or water. Typical yields are 20-30 μg for high-copy number plasmid or 3-10 μg for low-copy number plasmid from 4 ml of cultured bacterial cells. This plasmid kit does not require DNA phenol extraction or alcohol precipitation and the entire procedure can be completed within 15 minutes.
Video 1 .A step by step demonstrational video of the High-Speed Plasmid Mini Kit. Plasmid DNA from a 4 ml overnight E. coli (DH5α) culture, containing plasmid pBluescript (A600 > 2 U/ml) is purified using the High-Speed Plasmid Mini Kit.
Components
PD1, PD2, PD3 Buffers
W1 Buffer
Wash Buffer
Elution Buffer (10 mM Tris-HCl, pH8.5 at 25°C)
RNase A
PD Columns
2 ml Collection Tubes
Quality Control
The quality of the High-Speed Plasmid Mini Kit is tested on a lot-to-lot basis according to Geneaid’s ISO-certified quality management system. Plasmid DNA from a 4 ml overnight E. coli (DH5α) culture, containing plasmid pBluescript (A600 > 2 U/ml) is purified using the High-Speed Plasmid Mini Kit. Following the purification process, a yield of more than 20 μg is obtained and the A260/A280 ratio is between 1.8-2.0. The purified plasmid (1 µg) is used in EcoRI digestion, and checked by electrophoresis.