NucleoSpin Blood, Mini Kit für DNA aus Blut
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NucleoSpin Blood ist ein Spin-Säulen-basiertes Kit für den manuellen Gebrauch. Es ist für die Isolierung von DNA aus Vollblut (stabilisiert mit EDTA, Citrat oder Heparin), Serum, Plasma und anderen Körperflüssigkeiten vorgesehen. Neben Spin-Säulen und Puffern enthält es Proteinase K für eine effiziente Lyse des Blutes.
| Anwendung | Isolierung von DNA |
|---|---|
| Automatische Nutzung | Nein |
| CE-zertifiziert | Nein, nur für wissenschaftlichen Gebrauch |
| Elutionsvolumen | 60–200 µL |
| Format | Mini Präp |
| Fragmentgröße | 200 bp–ca. 50 kbp |
| Gefahrstoffe | Ja |
| Haltbarkeit (ab Produktion) | 27 Monat(e) |
| Handhabung | Zentrifugation |
| Lagertemperatur | 15–25 °C |
| Präparationszeit | 30 min/Präp |
| Probenmaterial | Blut, Blutplättchen, Buffy Coat, Körperflüssigkeiten, Plasma, Serum, Zellen |
| Probenmenge | < 5 x 10⁶ menschliche/tierische Zellen, kultivierte Zellen |
| Technologie | Silica-Membrantechnologie |
| Theoretische Bindungskapazität | 60 µg |
| Typische Ausbeute | 4–6 µg |
| Typische Downstream-Anwendung | Next Generation Sequenzierung, PCR, Southern Blotting, enzymatische Reaktionen |
| Typische Konzentration | 40–100 ng/µL |
| Typische Reinheit A260/A280 | 1,6–1,9 |
| Verkaufseinheit | 10 Präp(s), 50 Präp(s), 250 Präp(s) |
| Warenzeichen | NucleoSpin |
| Ziel | DNA |
NucleoSpin Blood
- Ready-to-use DNA in less than 30 min
- Complete removal of PCR inhibitors
- Suitable for whole blood, treated with citrate, EDTA, heparin, CPDA
- Consistently high DNA yields
NucleoSpin Blood - Procedure
NucleoSpin Blood kits are designed for rapid purification of genomic DNA from whole blood, serum, plasma, and body fluids (amniotic fluid, liquor etc.). Animal or human blood samples, fresh or frozen material, can be processed conveniently.
The NucleoSpin Blood method is based on a specially treated silica membrane with a high binding capacity for nucleic acids. The obtained DNA can be used directly for PCR (e.g., HLA typing), Southern blotting, or any kind of enzymatic reactions.
The kit allows the isolation of 4–6 μg of pure genomic DNA from 200 μL whole blood with an A₂₆₀/A₂₈₀ ratio between 1.60 and 1.90 and a typical concentration of 40–60 ng per μL. Due to the high binding capacity of the NucleoSpin Blood membrane, yields of up to 60 μg DNA are possible (from starting materials like buffy coat or lymphocyte-enriched samples).
With the NucleoSpin Blood method, genomic DNA is prepared from whole blood. Lysis is achieved by incubation of whole blood in a solution containing large amounts of chaotropic ions in the presence of proteinase K at 70 °C. Appropriate conditions for binding of DNA to the silica membrane of the NucleoSpin Blood columns are created by addition of ethanol to the lysate.
Superior purification with the NucleoSpin Blood kit
DNA was isolated from human blood samples (n = 3) using the NucleoSpin Blood kit and competitor kits from T, AJ and Q (light grey bars for A₂₆₀/A₂₈₀ and dark grey bars for A₂₆₀/A₂₃₀ ratios). The purity was determined by UV-spectrometry resulting in an average A₂₆₀/A₂₈₀ value for the NucleoSpin Blood kit of 1.89 ± 0.03 (dark blue bar) and an average A₂₆₀/A₂₃₀ value (light blue bar) of 1.88 ± 0.03.
Competitive sensitivity measured by qPCR
DNA was extracted from human blood samples the with NucleoSpin Blood kit (dark blue bar) and the competitor kits from T, AJ and Q (grey bars). Samples were analyzed in triplicate by qPCR for β-globin (268 bp). With an average amplification cycle of 23.1 the results demonstrate the competitive performance and reliably high quality of DNA extraction with the NucleoSpin Blood kit.
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