Introduction
HiPure Plasmid EF Maxi Kit Features:
- Technology Integration:
- The kit integrates HiPure technology for plasmid purification with Magen’s innovative Endotoxin Removal Technology (ETR).
- Endotoxin Removal:
- Specifically designed to achieve low levels of endotoxins in the purified plasmid DNA, enhancing its suitability for sensitive applications.
- Application Suitability:
- Ideal for use in eukaryotic transfection and various in vitro experiments due to the high purity and low endotoxin levels of the DNA.
- Capacity and Yield:
- Capable of purifying up to 1000 µg of high copy number plasmid DNA or 200 µg of low copy number plasmid DNA.
- Culture Volume Processing:
- Efficient purification from up to 150 mL of an overnight bacterial culture.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation of up to 1.5 mg endotoxin-free plasmid DNA from 200ml bacterial culture |
| Applications | Cell transfection, animal injection, etc. |
| Purification method | Maxi spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Conventional plasmid, large plasmid |
| Sample amount | High copy plasmid: 100-150ml culture medium
Low copy plasmid: 150-200ml culture medium |
| Yield | 100-1500μg |
| Elution volume | ≥0.8ml |
| Time per run | ≤60 minutes |
| Liquid carrying volume per column | 20ml |
| Binding yield of column | 1 mg |
Principle
HiPure Plasmid Procedure:
- Alkaline Lysis and DNA Adsorption:
- Based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt.
- The unique silica membrane in the kit replaces glass or silica slurries for plasmid DNA minipreps.
- Three Basic Steps:
- Step 1: Preparation and clearing of a bacterial lysate using the alkaline method.
- Step 2: Transfer of the supernatant to a column for DNA binding.
- Step 3: Washing to remove proteins and other impurities, followed by elution of nucleic acid with low-salt buffer (10 mm Tris, pH 9.0, 0.5 mm 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 1.5 mg plasmid can be binded in one column
- 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 | P115602 | P115603 |
| Purification Times | 10 Preps | 50 Preps |
| RNase A | 30 mg | 150 mg |
| Buffer P1 | 100 ml | 500 ml |
| Buffer P2 | 100 ml | 500 ml |
| Buffer LN3 | 50 ml | 250 ml |
| Buffer PW1 | 33 ml | 180 ml |
| Buffer PW2 | 20 ml | 100 ml |
| Elution Buffer | 15 ml | 120 ml |
| Buffer CL | 33 ml | 180 ml |
| HiPure DNA Maxi Columns C | 10 | 50 |
| Lysate Clear Midi Syringe | 10 | 50 |
| 50 ml Collection Tubes, C | 20 | 100 |
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.
- Buffer Precipitates:
- If any precipitates form in the buffers, warm at 37°C to dissolve.
- Specific Storage for RNase A and Buffer P1:
- RNase A and Buffer P1 are 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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