Introduction
The HiPure FastFilter ADN plasmidique Maxi Kits combine the power of HiPure technology with the time-tested consistency of alkaline-FDS lysis of bacterial cells to deliver high-quality DNA. HiPure DNA columns facilitate the binding, washing and elution steps thus enabling multiple samples to be simultaneously processed. This system also includes a special filter cartridge which replaces the centrifugation step following alkaline lysis. Plasmid DNA purified by this system is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and other manipulations. Jusqu'à 1000 µg high copy number plasmid DNA or 50-500 µg low copy number plasmid DNA can be purified from 200mL overnight culture.
Détails
Caractéristiques
| Caractéristiques | Caractéristiques |
| Fonctions principales | Isolement jusqu'à 1 mg d'ADN plasmidique à partir de 200 ml de culture bactérienne |
| Applications | Enzyme digestion, séquençage, RAP and labeling, etc.. |
| Méthode de purification | Maxi colonne de rotation |
| Technologie de purification | Technologie de la silice |
| Méthode de processus | Manuel (centrifugation ou vide) |
| Échantillon type | Conventional plasmid, plasmid less than 30KB |
| Montant de l'échantillon | 100-200ml |
| Rendement | 0.4-1mg |
| Volume d'élution | ≥500μl |
| Temps par course | ≤60 minutes |
| Volume de liquide transporté par colonne | 20ml |
| Rendement de liaison de la colonne | 1mg |
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 PCR, etc..
- Rapide – it takes only 15-60 minutes to complete the isolation
- Économie – high cost performance
Contenu du kit
| Contenu | P101402 | P101403 |
| Temps de purification | 10 Préparations | 50 Préparations |
| RNase A | 20 mg | 40 mg |
| Buffer P1 | 140 ml | 2 X 350 ml |
| Buffer P2 | 140 ml | 2 X 350 ml |
| Buffer LEN3 | 70 ml | 350 ml |
| Buffer GBT | 120 ml | 550 ml |
| Buffer PW1 | 60 ml | 300 ml |
| Buffer PW2* | 50 ml | 4 X 100 ml |
| Tampon d'élution | 20 ml | 120 ml |
| HiPure DNA Maxi Columns III | 10 | 50 |
| Lysate Clear Maxi Syringe | 10 | 50 |
| 50 Tubes de prélèvement ml | 20 | 100 |
Stockage et stabilité
The kit components can be stored dry at room temperature (15-25°C) et sont stables pendant au moins 18 mois dans ces 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.
Guide d'achat
| Nom | CAT NO | Montant de l'échantillon | Column type | Elutio volume | Rendement | Temps par course | Centrifuge requirements |
| Mini kit plasmidique HiPure | P1001 | 1-5ml | 1.5ml column | 30-50µl | 5–35µg/5ml | ≤ 30 minutes | small centrifuge |
| HiPure Plasmide Mini Kit II | P1002 | 5-15ml | 1.5ml column | 75-100µl | 20-80µg/15ml | ≤ 30 minutes | small centrifuge |
| HiPure Plasmide Plus 96 Trousse | P1006 | 1-1.5ml | 96 bien plaque | 70-150µl | 1-15μg/1ml | ≤60 minutes | 96 well plate centrifuge |
| Kit Midi Plasmide HiPure Fastfilter | P1013 | 30-50ml | 15ml column | 0.3-0.8ml | 50–250µg/50ml | ≤60 minutes | 15ml centrifuge |
| Kit Maxi Plasmide HiPure Fastfilter | P1014 | 100-200ml | 50ml column | 0.5-1.5ml | 0.4–1mg/200ml | ≤60 minutes | 50ml centrifuge |
Commentaires
Il n'y a pas encore de critiques.