Einführung
The HiPure Midi system provides a fast, simple, and cost-effective Plasmid-DNA 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.
Einzelheiten
Spezifikationen
| Merkmale | Spezifikationen |
| Hauptfunktionen | Isolation up to 500µg endotoxin-free plasmid DNA from 25-50ml bacterial culture |
| Anwendungen | Cell transfection, animal injection, usw. |
| Reinigungsmethode | Midi Spin-Säule |
| Reinigungstechnologie | Silica-Technologie |
| Prozessmethode | Handbuch (Zentrifugation oder Vakuum) |
| Beispielstyp | Conventional plasmid, plasmid less than 30KB |
| Probenmenge | 25-50ml |
| Ertrag | 10-500μg |
| Elutionsvolumen | ≥0.4ml |
| Zeit pro Lauf | ≤ 60 Minuten |
| Flüssigkeitstransportvolumen pro Säule | 4ml |
| Bindungsausbeute der Säule | 250μg |
Prinzip
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. Nach dem Waschen von Proteinen und anderen Verunreinigungen, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9,0, 0.5mm EDTA).
Vorteile
- Hohe Reinheit – purified plasmid can be directly used in sequencing, enzyme digestion and PCR, usw.
- Schnell – silica gel column purification is much faster than ion exchange
- Hohe Ausbeute – 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
Inhalt des Kits
| Inhalt | P123102 | P123102B |
| Reinigungszeiten | 50 Vorbereitungen | 50 Vorbereitungen |
| 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 |
| Elutionspuffer | 30 ml | 30 ml |
| MaxPure EF Mini Column | 50 | 50 |
| 2 ml-Sammelröhrchen | 50 | 50 |
| Lysate Clear Midi Syringe | 50 | 50 |
| Extend Tubes | 50 | 50 |
| 50ml Centrifuge Tubes | 50 | |
| Support Tubes | 50 |
Lagerung und Stabilität
The kit components can be stored dry at room temperature (15-25°C) und sind mindestens stabil 18 Monate unter diesen Bedingungen. Falls sich in den Puffern Ausfällungen bilden, Zum Auflösen auf 37 °C erwärmen. After addition of RNase A and optional LyseBlue reagent, Buffer P1 is stable for 6 months when stored at 2–8°C.
Kaufratgeber
| Name | KATZE NEIN | Probenmenge | Spaltentyp | Elutionsvolumen | Ertrag | Zeit pro Lauf | Centrifuge requirements |
| Hipure Plasmid EF Mini Kit | P1154 | 10-15ml | 1.5ml-Säule | ≥75μl | 10–70µg/15ml | ≤ 40 Minuten | small centrifuge |
| Hipure Plasmid EF 96 Bausatz | P1157 | 1-5ml | 96 Brunnenplatte | ≥75μl | 30µg/1ml | ≤ 60 Minuten | 96 well plate centrifuge |
| Hipure Plasmid EF MIDI Kit | P1231 | 30-50ml | 15ml-Säule | ≥0.4ml | 10-500μg/50ml | ≤ 60 Minuten | 15ml centrifuge |
| Hipure Plasmid EF Maxi Kit | P1156 | 100-200ml | 50ml-Säule | ≥0.8ml | 100–1500µg/200ml | ≤ 60 Minuten | 50ml centrifuge |
| Hipure Plasmid EF Mega Kit | P1116 | 500ml | 50ml-Säule | ≥0.8ml | 10mg/500ml | ≤ 70 Minuten | 50ml centrifuge |
Rezensionen
Es gibt noch keine Rezensionen.