Introduction
The HiPure Mini system provides a fast, simple, and cost-effective plasmid DNA miniprep method for routine molecular biology laboratory applications. HiPure Plasmid Mini Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of Tris buffer or water.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation up to 70μg plasmid DNA from 5-15ml bacterial culture |
| Applications | Enzyme digestion, sequencing, PCR, cloning, etc. |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Conventional plasmid, plasmid less than 30KB |
| Sample amount | High copy plasmid: 1-5ml culture medium Low copy plasmid : 5-10ml culture medium |
| Yield | 20-80µg |
| Elution volume | ≥75μl |
| Time per run | Complete 1-24 samples in 30 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 70µg |
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: Preparation and 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 2 minutes to obtain supernatant by optimized solution (10 minutes for other brands)
- High yield – up to 70µg plasmid can be binded in one column
Kit Contents
| Contents | P100202 | P100203 |
| Purification Times | 50 Preps | 250 Preps |
| RNase A | 5 mg | 20 mg |
| Buffer P1 | 30 ml | 140 ml |
| Buffer P2 | 30 ml | 140 ml |
| Buffer P3 | 40 ml | 200 ml |
| Buffer PW1 | 30 ml | 140 ml |
| Buffer PW2* | 12 ml | 50 ml |
| Elution Buffer | 15 ml | 30 ml |
| HiPure DNA Mini Columns III | 50 | 250 ml |
| 2 ml Collection Tubes | 50 | 250 ml |
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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