High-Speed Plasmid Mini Kit
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.
Advantages (Cat. # PD100, PD300)
- Purify plasmid DNA or cosmid DNA within 15 minutes!
- High Yield: up to 30 μg of pure plasmid DNA or cosmid DNA
- Sample Volume: 1-5 ml of bacterial cells
- Plasmid glass fiber spin column (red membrane optimized for plasmd DNA purification)
- Storage: dry at room temperature (15-25ºC) for up to 1 year
Applications
Restriction Enzyme Digestion, Library Screening, Ligation, PCR, Transformation and Sequencing Reactions
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 Plasmd 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 EcoR I digestion, and checked by electrophoresis.
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Figure 1. Plasmid DNA was extracted using both the High-Speed Plasmid Kit (lane 3,4) and the equivalent competitor's plasmid kit (lane 1,2). The purified supercoiled plasmid DNA [1.5 ml overnight E. coli (DH5α) culture, containing a 3 kb plasmid pBluescript (A600 > 2 U/ml)], from a 50 µl eluate was used in EcoRI digestion and analyzed by electrophoresis on a 1% agarose gel.
M = Geneaid 1 Kb DNA Ladder
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Figure 2. Sequencing data of (pBluescript) plasmid DNA purified using the High-Speed Plasmid Mini Kit
Click image to expand.
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Figure 3. A variety of plasmid and cosmid DNA was purified using the High-Speed Plasmid Mini Kit. Plasmid pBluescript II (lane 1), pUC18 (lane 2), pBR 322 (lane 3), pGEM with 1 kb insert (lane 4), pET 43.1 with 2.7 kb insert (lane 5) and cosmid (50 kb, lane 6) DNA from 50 µl eluates was used in EcoRI digestion and analyzed by electrophoresis on a 1% agarose gel.
M1 = Geneaid 1 Kb DNA Ladder, M2 = Lambda-Hind III
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| 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. |
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