Introduction
The HiPure Fastfilter Plasmid DNA Midi Kits combine the power of HiPure technology with the time-tested consistency of alkaline-SDS lysis of bacterial cells to deliver high-quality DNA. HiPure DNA columns facilitate the binding, washing and elution steps thus enabling multiple samples to be simultaneously processed. This system also includes aspecial filter cartridge which replaces the centrifugation step following alkaline lysis. Plasmid DNA purified by this system is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and other manipulations. Up to 250 µg high copy number plasmid DNA or 10-75 µg low copy number plasmid DNA can be purified from 15-50 mL overnight culture.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation up to 250μg plasmid DNA from 30-50ml bacterial culture |
| Applications | Enzyme digestion, sequencing, PCR and labeling, etc. |
| Purification method | Midi spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Conventional plasmid, plasmid less than 30KB |
| Sample amount | 30-50ml |
| Yield | 50-250µg |
| Elution volume | ≥300μl |
| Time per run | ≤60 minutes |
| Liquid carrying volume per column | 4ml |
| Binding yield of column | 250µg |
Principle
The HiPure Plasmid procedure is based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt. The unique silica membrane used in the kit completely replaces glass or silica slurries for plasmid DNA minipreps. The procedure consists of 3 basic steps: Preparationand clearing of a bacterial lysate by alkaline method，then transfer the supernatant to column to bind DNA. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).
Advantages
- High purity – purified plasmid can be directly used in sequencing, enzyme digestion and PCR, etc.
- Fast – it takes only 15-60 minutes to complete the isolation
- High yield – up to 70µg plasmid can be binded in one column
Kit Contents
| Contents | P101302 | P101303 |
| Purification Times | 25 Preps | 100 Preps |
| RNase A | 10 mg | 40 mg |
| Buffer P1 | 80 ml | 300 ml |
| Buffer P2 | 80 ml | 300 ml |
| Buffer LEN3 | 40 ml | 150 ml |
| Buffer GBT | 60 ml | 250 ml |
| Buffer PW1 | 60 ml | 250 ml |
| Buffer PW2 | 20 ml | 100 ml |
| Elution Buffer | 30 ml | 120 ml |
| HiPure DNA Midi Columns III | 25 | 100 |
| Lysate Clear Midi Syringe | 25 | 100 |
| 15 ml Collection Tubes | 50 | 200 |
Storage and Stability
The kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve. After addition of RNase A，Buffer P1 is stable for 6 months when stored at 2-8°C.
Purchase Guide
| Name | CAT NO | Sample amount | Column type | Elutio volume | Yield | Time per run | Centrifuge requirements |
| HiPure Plasmid Mini Kit | P1001 | 1-5ml | 1.5ml column | 30-50μl | 5–35µg/5ml | ≤30 minutes | small centrifuge |
| HiPure Plasmid Mini Kit II | P1002 | 5-15ml | 1.5ml column | 75-100μl | 20-80µg/15ml | ≤30 minutes | small centrifuge |
| HiPure Plasmid Plus 96 Kit | P1006 | 1-1.5ml | 96 well plate | 70-150μl | 1-15μg/1ml | ≤60 minutes | 96 well plate centrifuge |
| HiPure Fastfilter Plasmid Midi Kit | P1013 | 30-50ml | 15ml column | 0.3-0.8ml | 50–250µg/50ml | ≤60 minutes | 15ml centrifuge |
| HiPure Fastfilter Plasmid Maxi Kit | P1014 | 100-200ml | 50ml column | 0.5-1.5ml | 0.4–1mg/200ml | ≤60 minutes | 50ml centrifuge |
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