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Flexible magnetic bead based isolation of DNA from formalin-fixed paraffin-embedded samples
| Application | Isolation of DNA |
| Selling unit | 96 Prep(s), 384 Prep(s) |
| Target | DNA |
| CE certified | No, research use only |
| Technology | Magnetic bead technology |
| Brand | NucleoMag |
| Format | Magnetic beads |
| Handling | Magnetic separation |
| Lysate clarification | Centrifugation |
| 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 | ≤ 5 mg tissue, < 15 mg paraffin |
| Fragment size | 300 bp–approx. 50 kbp |
| Theoretical binding capacity | 0.4 µg/µL beads |
| Elution volume | > 25 µL |
| Preparation time | 40–120 min/96 preps (excl. lysis) |
| Typical downstream application | PCR |
| Storage temperature | 15–25 °C / 59–77 °F |
| Shelf life (from production) | 24 Month(s) |
| Hazardous material | Yes |
NucleoMag DNA FFPE kit is designed for high-throughput DNA purification from formalin-fixed, paraffin-embedded samples.
The NucleoMag 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.
The procedure is based on the reversible adsorption of nucleic acids to paramagnetic beads under appropriate buffer conditions. After magnetic separation, the paramagnetic beads are washed to remove contaminants and salts using two different wash buffers. Pure genomic DNA is finally eluted under low ionic strength conditions in a slightly alkaline elution buffer.
DNA was isolated from formalin-fixated and paraffin-embedded mouse sections (thickness 20 μm each) with the NucleoMag DNA FFPE kit. Dimensions of each section: brain 50 mm2 ; liver 70 mm2 ; lung 40 mm2 ; kidney 40 mm2 . Extraction of 1 slice / prep (4 preparations for each organ) resulted in the indicated total amounts of DNA (dark blue bars) measured via optical density. The qPCR analysis (orange squares) was performed using Thermo Scientific™ Maxima SYBR Green qPCR Master Mix.
DNA was isolated from mouse FFPE samples (n = 4; approximate 10 mg paraffin each) using the NucleoMag® DNA FFPE kit in an automated manner on an epMotion® 5075t system (dark blue bars) or manually (grey bars). A subsequent qPCR analysis was performed with a Taqman® Probe for a GADPH amplicon using the SensiFastTM Probe Lo-ROX kit from Bioline on an Applied Biosystems® 7500 Real-Time PCR System. The results demonstrate a reliable performance of the established, automated method.
DNA was isolated from various mouse FFPE samples (n = 4; approximate section size muscle: 1 mm²; liver: 12 mm²; esophagus: 3 mm²; lung: 5 mm²; kidney: 8 mm²; brain: 4,5 mm²) using the NucleoMag® DNA FFPE kit on an epMotion® 5075t system. The total yield was determined by UV spectrometry (dark blue bars). A subsequent qPCR analysis was performed with a Taqman® Probe for a GADPH amplicon using the SensiFastTM Probe Lo-ROX kit from Bioline on an Applied Biosystems® 7500 Real-Time PCR System (orange squares). The results demonstrate a reliable qPCR-performance for all tested mouse FFPE samples.
MACHEREY-NAGEL delivers a ready to go solution for your automated FFPE sample handling. We adapted the NucleoMag DNA FFPE procedure on the epMotion 5075t system to facilitate your DNA extraction workflow.
MACHEREY-NAGEL delivers a ready to go solution for your high throughput FFPE sample processing. We adapted the NucleoMag DNA FFPE procedure on the KingFisher™ instruments to speed up your nucleic acid purification workflow.
DNA was isolated from formalin-fixated and paraffin-embedded human sections (thickness 20 μM each). Dimensions of each section: spleen 150 mm2 ; stomach 150 mm2 . Extraction of 1 slice/prep (4 preparations for each organ) resulted in the indicated total amounts of DNA (dark blue bars) and qPCR values (orange squares). Analysis was performed using the Quantifiler™ Human DNA Quantification Kit from Thermo Scientific™.
