NucleoSpin 96 DNA FFPE, 96-well Kit für DNA aus FFPE Proben
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Isolation of DNA from formalin-fixed paraffin-embedded samples in proven 96‑well plate format
| Anwendung | Isolierung von DNA |
|---|---|
| Automatische Nutzung | Ja |
| CE-zertifiziert | Nein, nur für wissenschaftlichen Gebrauch |
| Elutionsvolumen | 100 µL |
| Format | 96-Well-Platte |
| Fragmentgröße | 50 bp–ca. 5 kbp |
| Gefahrstoffe | Ja |
| Haltbarkeit (ab Produktion) | 24 Monat(e) |
| Handhabung | Zentrifugation, Vakuum |
| Kompatibilität der Automatisierung | Kompatibel mit den meisten offenen Roboterplattformen. Bitte besuchen Sie www.mn-net.com/automation oder wenden Sie sich an unseren technischen Support für weitere Informationen. |
| Lagertemperatur | 15–25 °C |
| Präparationszeit | 60 min/Platte (ohne Lyse) |
| Probenmaterial | FFPE, Fixierte Proben, Gewebeschnitte |
| Probenmenge | < 10 mg Gewebe, < 15 mg Paraffin |
| Technologie | Silica-Membrantechnologie |
| Theoretische Bindungskapazität | 20 µg |
| Typische Ausbeute | Abhängig von der Menge und Qualität der Probe |
| Typische Downstream-Anwendung | PCR |
| Typische Reinheit A260/A280 | Stark abhängig von der Probenqualität |
| Verkaufseinheit | 96 Präp(s), 384 Präp(s) |
| Warenzeichen | NucleoSpin |
| Ziel | DNA |
NucleoSpin96 DNA FFPE
- Blue colored Paraffin Dissolver (patented technology) for easy paraffin removal
- Decrosslinking buffer to eliminate formalin crosslinking
- High quality of DNA for improved performance in downstream applications (e.g., PCR)
- Processing under vacuum or by centrifugation, suitable for manual and automated processing
NucleoSpin96 DNA FFPE procedure
NucleoSpin96 DNA FFPE kit is designed for high-throughput DNA purification from formalin-fixed, paraffin-embedded samples. The NucleoSpin96 DNA FFPE kit contains the odorless Paraffin Dissolver (patent pending) which allows effective lysis in a convenient two-phase system. First, after dissolving of paraffin from FFPE sections with Paraffin Dissolver, the tissue sample is digested by Proteinase K to solubilize the fixed tissue and release DNA into solution. Subsequently, heat incubation with a specially designed buffer effectively eliminates crosslinks from the previously released DNA.
Appropriate conditions for binding of DNA to the silica membrane in the NucleoSpin DNA FFPE Binding Plate are created by addition of large amounts of chaotropic salt and ethanol to the lysate. The binding process is reversible and specific to nucleic acids. While DNA is kept on the silica membrane, contaminations are removed by washing with two different wash buffers. Pure genomic DNA is finally eluted under low ionic strength conditions in a slightly alkaline elution buffer.
Reliable DNA yield
DNA was purified from seven human ileum as well as human stomach samples and quantified with Quantifiler Human DNA Quantification Kit. A DNA yield of approximately 0.3 μg from ileum and 1.7 μg from stomach could be obtained.
Consistent PCR performance
Genomic DNA was isolated with the NucleoSpin 96 DNA FFPE kit from mouse liver, lung, muscle, and kidney samples (eight samples each). The purified DNA was quantified with MaximaTM SYBR Green qPCR Master Mix (amplicon size 100 bp) resulting in reliable CT values indicating consistent DNA yields.
Genomic DNA was isolated with the NucleoSpin 96 DNA FFPE kit from mouse liver, lung, muscle, and kidney samples (eight samples each). Agarose gel electrophoresis of PCR product from mouse liver samples demonstrates reliable PCR results (lane 2–9).
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