The HiYield Plasmid Mini Kit is designed for rapid isolation of plasmid DNA from 1-4 ml of cultured bacterial cells. Modified alkaline lysis method and RNase treatment are used to obtain clear cell lysate with minimal genomic DNA and RNA contaminants. In the presence of chaotropic salt, plasmid DNA in the lysate binds to the glass fiber matrix of the spin column. Contaminants are removed with a Wash Buffer (containing ethanol) and the purified plasmid DNA is eluted by a low salt Elution Buffer or TE. 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. DNA phenol extraction or alcohol precipitation is not required and the entire procedure can be completed within 30 minutes.
- Sample :
1-4 ml of bacterial cells - Format :
spin column (centrifuge) - Yield :
up to 30 μg of plasmid/cosmid DNA
Restriction enzyme digestion, library screening, ligation, PCR, transformation/sequencing reactions
The quality of the HiYield Plasmid Mini Kit is tested on a lot-to-lot basis, by isolating plasmid DNA from a 4 ml overnight E. coli (DH5α) culture, containing plasmid pBluescript (A600 > 2 U/ml). Following the purification process, a yield of more than 20 μg is expected and the ratio of A260/A280 is between 1.7-1.9. The purified plasmid (1 μg) is used in EcoR I digestion, and checked by electrophoresis.
| PD1Buffer | 25 ml |
| PD2 Buffer | 25 ml |
| PD3 Buffer | 45 ml |
| W1 Buffer | 45 ml |
| Wash Buffer | 25 ml |
| Elution Buffer | 6 ml |
| RNase A (50 mg/ml) | 50 μl |
| PD Column | 100 pcs |
| 2 ml Collection Tube | 100 pcs |
| PD1Buffer | 65 ml |
| PD2 Buffer | 75 ml |
| PD3 Buffer | 100 ml |
| W1 Buffer | 130 ml |
| Wash Buffer | 50 ml |
| Elution Buffer | 30 ml |
| RNase A (50 mg/ml) | 130μl |
| PD Column | 300 pcs |
| 2 ml Collection Tube | 300 pcs |
