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 Purification de l'ADN.
- 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.
Détails
Caractéristiques
Caractéristiques | Caractéristiques |
Fonctions principales | Isolement jusqu'à 70 µg d'ADN plasmidique sans endotoxine à partir de 10 à 15 ml de culture bactérienne |
Applications | Cell transfection, animal injection, etc.. |
Méthode de purification | Mini colonne de rotation |
Technologie de purification | Technologie de la silice |
Méthode de processus | Manuel (centrifugation ou vide) |
Échantillon type | Conventional plasmid |
Montant de l'échantillon | High copy plasmid: 10ml culture medium
Low copy number plasmid : 10-15ml culture medium |
Rendement | 10-70µg |
Volume d'élution | ≥75μl |
Temps par course | ≤40 minutes |
Volume de liquide transporté par colonne | 800µl |
Rendement de liaison de la colonne | 70µg |
Principe
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. Après avoir lavé les protéines et autres impuretés, l'acide nucléique a finalement été élué avec un tampon à faible teneur en sel (10mmTris, pH9,0, 0.5mmEDTA).
Avantages
- Haute pureté – purified plasmid can be directly used in sequencing, enzyme digestion and RAP, etc..
- Rapide – silica gel column purification is much faster than ion exchange
- Haut rendement – 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
Contenu du kit
Contenu | P115402 | P115403 |
Temps de purification | 50 Préparations | 250 Préparations |
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 |
Tampon d'élution | 15 ml | 30 ml |
HiPure DNA Mini Columns III | 50 | 250 |
2 Tubes de prélèvement ml | 50 | 250 |
Stockage et stabilité
Instructions de stockage:
- General Storage:
- Kit components can be stored dry at room temperature (15–25°C).
- Stable pour au moins 18 mois dans ces 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.
Guide d'achat
Nom | CAT NO | Montant de l'échantillon | Column type | Volume d'élution | Rendement | Temps par course | Centrifuge requirements |
HiPure Plasmide EF Mini Kit | P1154 | 10-15ml | 1.5ml column | ≥75μl | 10–70µg/15ml | ≤40 minutes | small centrifuge |
HiPure Plasmide EF 96 Trousse | P1157 | 1-5ml | 96 bien plaque | ≥75μl | 30µg/1ml | ≤60 minutes | 96 well plate centrifuge |
Kit HiPure Plasmid EF Midi | 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 Méga Kit | P1116 | 500ml | 50ml column | ≥0.8ml | 10mg/500ml | ≤70 minutes | 50ml centrifuge |
Commentaires
Il n'y a pas encore de critiques.