Introduction
The HiPure Minipreps system provides a fast, simple, and cost-effective plasmid DNA miniprep method for routine molecular biology laboratory applications. HiPure Miniprep 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 30μg endotoxin-free plasmid DNA from 1-5ml bacterial culture using 96-well bind plate |
| Applications | Cell transfection, animal injection, etc. |
| Purification method | 96 well plate |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Conventional plasmid, plasmid less than 30KB |
| Sample amount | 1-5ml（x96） |
| Yield | 30μg |
| Elution volume | ≥75μl |
| Time per run | ≤60 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 – silica gel column purification is much faster than ion exchange
- Ultra low endotoxin – content of endotoxin is less than 0.05 EU/μg, which can be directly used for cell transfection and animal injection, etc.
Kit Contents
| Contents | P115701 | P115702 | P115703 |
| Purification Times | 1 x 96 Preps | 4 x 96 Preps | 20 x 96 Preps |
| RNase A | 10 mg | 20 mg | 100 mg |
| Buffer P1 | 30 ml | 120 ml | 550 ml |
| Buffer P2 | 30 ml | 120 ml | 550 ml |
| Buffer LEN3 | 15 ml | 60 ml | 300 ml |
| Buffer LN4 | 80 ml | 270 ml | 2 x 700 ml |
| Buffer LN5 | 40 ml | 220 ml | 1100 ml |
| Buffer PW1 | 40 ml | 220 ml | 1100 ml |
| Buffer PW2 | 25 ml | 100 ml | 2 x 200 ml |
| Elution Buffer | 30 ml | 60 ml | 250 ml |
| Lysate Clear Plate | 1 | 4 | 20 |
| HiPure DNA Plate | 1 | 4 | 20 |
| 1.6ml Collection Plate | 2 | 8 | 40 |
| 0.5ml Collection Plate | 1 | 4 | 20 |
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 | 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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