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-RAP, Transfert du Nord, poly A+ RNA (ARNm) purification, nuclease protection, and in vitro translation.
Détails
Caractéristiques
| Caractéristiques | Caractéristiques |
| Fonctions principales | Isolation of total RNA from Paxgene RNA Tubes |
| Applications | RT-PCR, qPCR, Northern hybridization, NGS, nucleic acid protection, in vitro translation |
| Méthode de purification | Mini spin column |
| Technologie de purification | Technologie de la silice, DNA filtration technology |
| Méthode de processus | Manuel (centrifugation ou vide) |
| Échantillon type | Blood in the preservation tube |
| Montant de l'échantillon | 2.5ml |
| Volume d'élution | ≥30µl |
| Volume de liquide transporté par colonne | 800µl |
| Rendement de liaison de la colonne | 100µg |
Principe
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.
Avantages
- High quality – high purity total RNA can be directly used in various sensitive downstream applications
- Rapide – isolation of several samples can be completed in 40 minutes by using the column purification method
- Sécurité – no phenol-chloroform extraction is required
- Sensitive – direct lysis of blood, plasma, and other samples without separation of leukocytes
Contenu du kit
| Contenu | R416802 | R416803 |
| Temps de purification | 10 Préparations | 50 Préparations |
| HiPure RNA Mini Columns I | 10 | 50 |
| gDNA Filter Mini Columns | 10 | 50 |
| 2Tubes de prélèvement ml | 30 | 150 |
| RNase Free Water | 60 ml | 250 ml |
| Buffer MBR1 | 10 ml | 30 ml |
| Buffer MBR2 | 5 ml | 15 ml |
| Buffer RW1 | 15 ml | 60 ml |
| Buffer RW2* | 6 ml | 20 ml |
| Protéinase K | 12 mg | 50 mg |
| Tampon de dissolution de protéase | 1.8 ml | 5 ml |
| DNase I | 120 µl | 600 µl |
| DNase Buffer | 6 ml | 30 ml |
Stockage et stabilité
Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. Cependant, stockage à court terme (DNase I up to 1 semaines, Proteinase K up to 8 semaines) à température ambiante (15–25°C) n'affecte pas leurs performances. Les composants restants du kit peuvent être conservés à température ambiante (15–25°C) et sont stables pendant au moins 18 mois dans ces conditions.
Données d'expérimentation
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