Introduction
The HiPure Midi system provides a fast, simple, and cost-effective plasmid DNA midiprep method for routine molecular biology laboratory applications. HiPure Midiprep 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 500µg endotoxin-free plasmid DNA from 25-50ml bacterial culture |
| Applications | Cell transfection, animal injection, 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 | 25-50ml |
| Yield | 10-500μg |
| Elution volume | ≥0.4ml |
| 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: 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 – silica gel column purification is much faster than ion exchange
- High yield – up to 250μg plasmid can be binded in one column
- Low endotoxin content – the obtained plasmid can be directly used in cell transfection and animal injection
Kit Contents
| Contents | P123102 | P123102B |
| Purification Times | 50 Preps | 50 Preps |
| RNase A | 30 mg | 30 mg |
| Buffer E1 | 150 ml | 150 ml |
| Buffer E2 | 150 ml | 150 ml |
| Buffer E3 | 150 ml | 150 ml |
| Buffer E4 | 150 ml | 150 ml |
| Buffer E5 | 150 ml | 150 ml |
| Buffer PW2* | 50 ml | 50 ml |
| Elution Buffer | 30 ml | 30 ml |
| MaxPure EF Mini Column | 50 | 50 |
| 2 ml Collection Tubes | 50 | 50 |
| Lysate Clear Midi Syringe | 50 | 50 |
| Extend Tubes | 50 | 50 |
| 50ml Centrifuge Tubes | 50 | |
| Support Tubes | 50 |
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 and optional LyseBlue reagent, Buffer P1 is stable for 6 months when stored at 2–8°C.
Purchase Guide
| Name | CAT NO | Sample amount | Column type | Elution volume | Yield | Time per run | Centrifuge requirements |
| HiPure Plasmid EF Mini Kit | P1154 | 10-15ml | 1.5ml column | ≥75μl | 10–70µg/15ml | ≤40 minutes | small centrifuge |
| HiPure Plasmid EF 96 Kit | P1157 | 1-5ml | 96 well plate | ≥75μl | 30µg/1ml | ≤60 minutes | 96 well plate centrifuge |
| HiPure Plasmid EF Midi Kit | P1231 | 30-50ml | 15ml column | ≥0.4ml | 10-500μg/50ml | ≤60 minutes | 15ml centrifuge |
| HiPure Plasmid EF Maxi Kit | P1156 | 100-200ml | 50ml column | ≥0.8ml | 100–1500µg/200ml | ≤60 minutes | 50ml centrifuge |
| HiPure Plasmid EF Mega Kit | P1116 | 500ml | 50ml column | ≥0.8ml | 10mg/500ml | ≤70 minutes | 50ml centrifuge |
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