Introduction
The HiPure Midi system provides a fast, simple, and cost-effective ADN plasmidique midiprep method for routine molecular biology laboratory applications. HiPure Midiprep Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of Tris buffer or water.
Détails
Caractéristiques
| Caractéristiques | Caractéristiques |
| Fonctions principales | Isolation up to 500µg endotoxin-free plasmid DNA from 25-50ml bacterial culture |
| Applications | Cell transfection, animal injection, etc.. |
| Méthode de purification | Midi 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 | 25-50ml |
| Rendement | 10-500µg |
| Volume d'élution | ≥0.4ml |
| Temps par course | ≤60 minutes |
| Volume de liquide transporté par colonne | 4ml |
| Rendement de liaison de la colonne | 250µg |
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 250μg 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 | P123102 | P123102B |
| Temps de purification | 50 Préparations | 50 Préparations |
| RNase A | 30 mg | 30 mg |
| Buffer E1 | 150 ml | 150 ml |
| Buffer E2 | 150 ml | 150 ml |
| Buffer E3 | 150 ml | 150 ml |
| Buffer E4 | 150 ml | 150 ml |
| Buffer E5 | 150 ml | 150 ml |
| Buffer PW2* | 50 ml | 50 ml |
| Tampon d'élution | 30 ml | 30 ml |
| MaxPure EF Mini Column | 50 | 50 |
| 2 Tubes de prélèvement ml | 50 | 50 |
| Lysate Clear Midi Syringe | 50 | 50 |
| Extend Tubes | 50 | 50 |
| 50ml Centrifuge Tubes | 50 | |
| Support Tubes | 50 |
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 and optional LyseBlue reagent, 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.