Einführung
The HiPure Fastfilter Plasmid-DNA Midi Kits combine the power of HiPure technology with the time-tested consistency of alkaline-Sicherheitsdatenblatt 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 aspecial 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. Bis zu 250 µg high copy number plasmid DNA or 10-75 µg low copy number plasmid DNA can be purified from 15-50 mL overnight culture.
Einzelheiten
Spezifikationen
| Merkmale | Spezifikationen |
| Hauptfunktionen | Isolation up to 250μg plasmid DNA from 30-50ml bacterial culture |
| Anwendungen | Enzyme digestion, Sequenzierung, PCR and labeling, usw. |
| Reinigungsmethode | Midi Spin-Säule |
| Reinigungstechnologie | Silica-Technologie |
| Prozessmethode | Handbuch (Zentrifugation oder Vakuum) |
| Beispielstyp | Conventional plasmid, plasmid less than 30KB |
| Probenmenge | 30-50ml |
| Ertrag | 50-250µg |
| Elutionsvolumen | ≥300μl |
| Zeit pro Lauf | ≤60 minutes |
| 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: Preparationand 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 – it takes only 15-60 minutes to complete the isolation
- Hohe Ausbeute – up to 70µg plasmid can be binded in one column
Inhalt des Kits
| Inhalt | P101302 | P101303 |
| Reinigungszeiten | 25 Vorbereitungen | 100 Vorbereitungen |
| RNase A | 10 mg | 40 mg |
| Buffer P1 | 80 ml | 300 ml |
| Buffer P2 | 80 ml | 300 ml |
| Buffer LEN3 | 40 ml | 150 ml |
| Buffer GBT | 60 ml | 250 ml |
| Buffer PW1 | 60 ml | 250 ml |
| Buffer PW2 | 20 ml | 100 ml |
| Elutionspuffer | 30 ml | 120 ml |
| HiPure DNA Midi Columns III | 25 | 100 |
| Lysate Clear Midi Syringe | 25 | 100 |
| 15 ml-Sammelröhrchen | 50 | 200 |
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，Buffer P1 is stable for 6 months when stored at 2-8°C.
Kaufratgeber
| Name | KATZE NEIN | Probenmenge | Spaltentyp | Elutio volume | Ertrag | Zeit pro Lauf | Centrifuge requirements |
| HiPure Plasmid Mini Kit | P1001 | 1-5ml | 1.5ml-Säule | 30-50μl | 5–35µg/5ml | ≤30 Minuten | small centrifuge |
| HiPure Plasmid Mini Kit II | P1002 | 5-15ml | 1.5ml-Säule | 75-100μl | 20-80µg/15ml | ≤30 Minuten | small centrifuge |
| HiPure Plasmid Plus 96 Bausatz | P1006 | 1-1.5ml | 96 Brunnenplatte | 70-150μl | 1-15μg/1ml | ≤60 minutes | 96 well plate centrifuge |
| HiPure Fastfilter Plasmid Midi Kit | P1013 | 30-50ml | 15ml-Säule | 0.3-0.8ml | 50–250µg/50ml | ≤60 minutes | 15ml centrifuge |
| HiPure Fastfilter Plasmid Maxi Kit | P1014 | 100-200ml | 50ml-Säule | 0.5-1.5ml | 0.4–1mg/200ml | ≤60 minutes | 50ml centrifuge |
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