Introduction
HiPure Plasmid EF Mini Kit Overview:
- Technology Integration:
- Combines HiPure technology with Magen’s innovative Endotoxin Removal Technology (ETR) to ensure high-quality plasmid DNA purification.
- Endotoxin Management:
- Features a specially formulated buffer designed to prevent endotoxins from binding to the HiPure matrix, significantly reducing endotoxin levels in the final DNA product.
- Application Suitability:
- Provides plasmid DNA ideal for eukaryotic transfection and in vitro experiments, especially where low endotoxin levels are critical.
- Impact of Endotoxin Contamination:
- Endotoxin contamination can drastically lower transfection efficiencies, particularly in endotoxin-sensitive cell lines.
- Clinical Relevance:
- In contexts such as gene therapy, controlling endotoxin levels is crucial due to the potential for serious adverse effects like fever, endotoxin shock syndrome, and negative impacts on in vitro transfection into immune cells.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation up to 70µg endotoxin-free plasmid DNA from 10-15ml bacterial culture |
| Applications | Cell transfection, animal injection, etc. |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Conventional plasmid |
| Sample amount | High copy plasmid: 10ml culture medium Low copy number plasmid : 10-15ml culture medium |
| Yield | 10-70μg |
| Elution volume | ≥75μl |
| Time per run | ≤40 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
- High yield – up to 70μg 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 | P115402 | P115403 |
| 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 LEN3 | 15 ml | 70 ml |
| Buffer LN4 | 50 ml | 250 ml |
| Buffer LN5 | 30 ml | 140 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 |
| 2 ml Collection Tubes | 50 | 250 |
Storage and Stability
Storage Instructions:
- General Storage:
- Kit components can be stored dry at room temperature (15–25°C).
- Stable for at least 18 months under these conditions.
- Handling Precipitates:
- If any precipitates form in the buffers, warm at 37°C to dissolve.
- Specific Storage for Buffer P1 with RNase A:
- 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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