Einführung
The HiPure Mini system provides a fast, simple, and cost-effective Plasmid-DNA miniprep method for routine molecular biology laboratory applications. HiPure Plasmid Mini 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 70μg plasmid DNA from 5-15ml bacterial culture |
| Anwendungen | Enzyme digestion, Sequenzierung, PCR, Klonen, usw. |
| Reinigungsmethode | Mini Spin-Säule |
| Reinigungstechnologie | Silica-Technologie |
| Prozessmethode | Handbuch (Zentrifugation oder Vakuum) |
| Beispielstyp | Conventional plasmid, plasmid less than 30KB |
| Probenmenge | High copy plasmid: 1-5ml culture medium
Low copy plasmid : 5-10ml culture medium |
| Ertrag | 20-80µg |
| Elutionsvolumen | ≥75μl |
| Zeit pro Lauf | Complete 1-24 samples in 30 Protokoll |
| Flüssigkeitstransportvolumen pro Säule | 800µl |
| Bindungsausbeute der Säule | 70µg |
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 – it takes only 2 minutes to obtain supernatant by optimized solution (10 minutes for other brands)
- Hohe Ausbeute – up to 70µg plasmid can be binded in one column
Inhalt des Kits
| Inhalt | P100202 | P100203 |
| Reinigungszeiten | 50 Vorbereitungen | 250 Vorbereitungen |
| RNase A | 5 mg | 20 mg |
| Buffer P1 | 30 ml | 140 ml |
| Buffer P2 | 30 ml | 140 ml |
| Buffer P3 | 40 ml | 200 ml |
| Buffer PW1 | 30 ml | 140 ml |
| Buffer PW2* | 12 ml | 50 ml |
| Elutionspuffer | 15 ml | 30 ml |
| HiPure DNA Mini Columns III | 50 | 250 ml |
| 2 ml-Sammelröhrchen | 50 | 250 ml |
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 Minuten | 96 well plate centrifuge |
| Hipure Fastfilter Plasmid Midi Kit | P1013 | 30-50ml | 15ml-Säule | 0.3-0.8ml | 50–250µg/50ml | ≤ 60 Minuten | 15ml centrifuge |
| Hipure Fastfilter Plasmid Maxi Kit | P1014 | 100-200ml | 50ml-Säule | 0.5-1.5ml | 0.4–1mg/200ml | ≤ 60 Minuten | 50ml centrifuge |
Rezensionen
Es gibt noch keine Rezensionen.