Introduction
The HiPure ADN plasmidique Mega 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. 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 10 mg high copy number plasmid DNA can be purified from 500 mL overnight culture.
Détails
Caractéristiques
Caractéristiques | Caractéristiques |
Fonctions principales | Isolement jusqu'à 10 mg d'ADN plasmidique sans endotoxine à partir de 500 ml de culture bactérienne |
Applications | Cell transfection, animal injection, etc.. |
Méthode de purification | Mega spin column |
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 | High copy plasmid: 500ml culture medium |
Yield | 10mg |
Volume d'élution | ≥0.8ml |
Temps par course | ≤70 minutes |
Volume de liquide transporté par colonne | 50ml |
Rendement de liaison de la colonne | 10mg |
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 10mg plasmid can be binded in one column
- Low endotoxin – the obtainedplasmid can be directly used for cell transfection and animal injection, etc..
Contenu du kit
Contenu | P111602 | P111603 |
Temps de purification | 10 Préparations | 50 Préparations |
RNase A | 60 mg | 2 X 150 mg |
Buffer E1 | 220 ml | 2 X 550 ml |
Buffer E2 | 220 ml | 2 X 550 ml |
Buffer E3 | 220 ml | 2 X 550 ml |
Buffer E4 | 220 ml | 2 X 550 ml |
Buffer E5 | 120 ml | 550 ml |
Buffer PW2 | 25 ml | 2 X 100 ml |
Tampon d'élution | 30 ml | 120 ml |
HiPure EF Maxi Columns | 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.
Guide d'achat
Nom | CAT NO | Montant de l'échantillon | Column type | Volume d'élution | Yield | 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 well plate | ≥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.