Introduction
The MagPure Plasmid purification system uses the paramagnetic bead technology for high-throughput preparation of high-copy or low-copy plasmid DNA from E. coli cells. This kit also can be used with fosmid and BAC vector-based constructs. The system uses alkaline lysis followed by a MagPure purification to differentially bind plasmid DNA to paramagnetic beads. While the DNA is bound to the beads, contaminants can be rinsed away using a simple washing procedure. Because MagPure uses magnetic separation technology, the protocol does not require vacuum filtration. This makes kit extremely amenable to automation. Plasmid DNA purified with this system is most commonly used in Sanger Sequencing and PCR amplification.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation up to 15μg endotoxin free plasmid DNA from 1-5ml bacterial culture |
| Applications | Enzyme digestion, sequencing, PCR and labeling, etc. |
| Purification technology | Magnetic beads technology |
| Process method | Manual or automatic |
| Sample type | Conventional plasmid, plasmid≤30KB |
| Sample amount | 1-5ml |
| Elution volume | ≥50μl |
| Time per run | ≤80 minutes |
Principle
This product is based on the purification method of high-binding magnetic particles. The sample is lysed and digested under the action of lysate and Lysozyme. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with a washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally, DNA was eluted with an Elution Buffer.
Advantages
- High purity – purified plasmid can be directly used in sequencing, enzyme digestion PCR, etc.
- Fast – it takes only 80 minutes to complete the isolation
- High yield – up to 15μg plasmid can be bound in one column
- Economic – excellent cost-effectiveness performance
Kit Contents
| Contents | P181402 | P181403 | P181404 |
| Purification Times | 100 Preps | 500 Preps | 5000 Preps |
| RNase A | 10 mg | 30 mg | 2 x 160 mg |
| Buffer P1 | 30 ml | 150 ml | 2 x 800 ml |
| Buffer P2 | 30 ml | 150 ml | 2 x 800 ml |
| Buffer LEN3 | 20 ml | 80 ml | 800 ml |
| Buffer LN4 | 90 ml | 400 ml | 4 x 980 ml |
| MagPure Particles | 3.5 ml | 17 ml | 3 x 60 ml |
Storage and Stability
RNase A and MagPure Particles should be stored at 2–8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15–25°C) does not affect its performance. The remaining kit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2–8°C, but in this case, buffers should be redissolved before use. Make sure that all buffers are at room temperature when used. If any precipitates form in the buffers, warm at 37℃ to dissolve. After the addition of RNase A, Buffer P1 is stable for 6 months when stored at
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