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Isolation of DNA from formalin-fixed paraffin-embedded samples in proven 96‑well plate format
| Application | Isolation of DNA |
| Selling unit | 96 Prep(s), 384 Prep(s) |
| Target | DNA |
| CE certified | No, research use only |
| Technology | Silica membrane technology |
| Brand | NucleoSpin |
| Format | 96-well plate |
| Handling | Centrifugation, vacuum |
| Automated use | Yes |
| Automation support | Compatible with most open robotic platforms. Please visit www.mn-net.com/automation or consult our technical support for further information. |
| Sample material | FFPE, Fixed samples, Tissue sections |
| Sample amount | ≤ 7 sections (10 µm) of 250 mm² total area, < 10 mg tissue, < 15 mg paraffin |
| Fragment size | 50 bp–approx. 5 kbp |
| Typical yield | Depending on amount and quality of sample |
| Theoretical binding capacity | 20 µg |
| Typical purity A260/A280 | Strongly depending on sample quality |
| Elution volume | 100 µL |
| Preparation time | 60 min/plate (excl. lysis) |
| Typical downstream application | PCR |
| Storage temperature | 15–25 °C / 59–77 °F |
| Shelf life (from production) | 24 Month(s) |
| Hazardous material | Yes |
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.
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.
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).
