The Kit provides fast purification of high-quality RNA from paxigene Blood RNA Tube using silica-membrane spin columns. RNA purified using the HiPure System is ready for applications such as RT-PCR, Northern Blot, poly A+ RNA (mRNA) Reinigung, nuclease protection, and in vitro translation.
Einzelheiten
Spezifikationen
| Merkmale | Spezifikationen |
| Hauptfunktionen | Isolation of total RNA from Paxgene RNA Tubes |
| Anwendungen | RT-PCR, qPCR, Nordhybridisierung, NGS, nucleic acid protection, in vitro translation |
| Reinigungsmethode | Mini Spin-Säule |
| Reinigungstechnologie | Silica-Technologie, DNA filtration technology |
| Prozessmethode | Handbuch (Zentrifugation oder Vakuum) |
| Beispielstyp | Blood in the preservation tube |
| Probenmenge | 2.5ml |
| Elutionsvolumen | ≥30μl |
| Flüssigkeitstransportvolumen pro Säule | 800µl |
| Bindungsausbeute der Säule | 100μg |
Prinzip
The Kit is for the purification of total RNA from 2.5 ml human whole blood collected in a PAXgene Blood RNA Tube. Purification begins with a centrifugation step to pellet nucleic acids in the PAXgene Blood RNA Tube. The pellet is washed and resuspended, and incubated in optimized buffers together with proteinase K. DNA wash The lysate is passed through a DNA Mini column. Ethanol is added to adjust binding conditions, and the lysate is applied to a column.RNA is selectively bound to the silica membrane as contaminants pass through. The remaining contaminants are removed in several efficient wash steps. Between the first and second wash steps, the membrane is treated with DNase I to remove trace amounts of bound DNA. After the wash steps, RNA is eluted in elution buffer and heat-denatured.
Vorteile
- Gute Qualität – high purity total RNA can be directly used in various sensitive downstream applications
- Schnell – isolation of several samples can be completed in 40 minutes by using the column purification method
- Safety – no phenol-chloroform extraction is required
- Sensitive – direct lysis of blood, Plasma, and other samples without separation of leukocytes
Inhalt des Kits
| Inhalt | R416802 | R416803 |
| Reinigungszeiten | 10 Vorbereitungen | 50 Vorbereitungen |
| HiPure RNA Mini Columns I | 10 | 50 |
| gDNA Filter Mini Columns | 10 | 50 |
| 2ml-Sammelröhrchen | 30 | 150 |
| RNase-freies Wasser | 60 ml | 250 ml |
| Buffer MBR1 | 10 ml | 30 ml |
| Buffer MBR2 | 5 ml | 15 ml |
| Puffer RW1 | 15 ml | 60 ml |
| Puffer RW2* | 6 ml | 20 ml |
| Proteinase K | 12 mg | 50 mg |
| Protease-Auflösungspuffer | 1.8 ml | 5 ml |
| DNase I | 120 µl | 600 µl |
| DNase Buffer | 6 ml | 30 ml |
Lagerung und Stabilität
Proteinase K sollte bei der Ankunft bei 2–8 °C gelagert werden. DNase I should be stored at -20°C. Jedoch, kurzfristige Lagerung (DNase I up to 1 Wochen, Proteinase K up to 8 Wochen) bei Raumtemperatur (15–25°C) hat keinen Einfluss auf ihre Leistung. Die restlichen Kit-Komponenten können bei Raumtemperatur gelagert werden (15–25°C) und sind mindestens stabil 18 Monate unter diesen Bedingungen.
Experimentdaten
Rezensionen
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