Figure 1. NucleoSpin 96 DNA FFPE procedure. The protocol can be performed using centrifugation or vacuum processing and can be automated on common liquid-handling platforms.
Product overview
NucleoSpin 96 DNA FFPE
The NucleoSpin 96 DNA FFPE kit is designed for high-throughput DNA purification from formalin-fixed, paraffin-embedded samples. Paraffin is first removed from the sample using odorless, xylene-free Paraffin Dissolver, which enables effective lysis in a convenient, two-phase system. Samples are then digested by Proteinase K to solubilize fixed tissues and release DNA into solution. Subsequently, crosslinks are removed via heat incubation in a specially designed buffer, and large amounts of chaotropic salt and ethanol are added to the lysate, providing conditions suitable for binding of DNA to the silica membrane in the NucleoSpin DNA FFPE Binding Plate. The binding process is reversible and specific to nucleic acids. While DNA is kept on the silica membrane, contaminants are removed by washing with two different wash buffers. Pure genomic DNA is eluted under low ionic strength conditions in a slightly alkaline elution buffer.
- Isolation of DNA from FFPE samples in a high-throughput, 96-well plate format
- Blue-colored, xylene-free Paraffin Dissolver for easy paraffin removal
- Decrosslinking buffer to eliminate formalin crosslinking
- High-quality DNA for improved performance in downstream applications
- Suitable for manual and automated processing under centrifugation or vacuum
Procedure
Reliable DNA yield
Figure 2. Reliable DNA yield with NucleoSpin 96 DNA FFPE. DNA was purified from seven FFPE samples derived from human ileum and stomach and quantified using the Quantifiler Human DNA Quantification Kit. Average DNA yields of 0.3 µg and 1.7 µg were obtained from the ileum and stomach, respectively.
Consistent PCR performance
Figure 3. Consistent PCR performance with NucleoSpin 96 DNA FFPE. Genomic DNA was isolated from mouse liver, lung, muscle, and kidney FFPE tissues (8 samples each) with the NucleoSpin 96 DNA FFPE kit. Panel A. Analysis by qPCR. Purified DNA was quantified using Maxima SYBR Green qPCR Master Mix (amplicon size: 100 bp). The similar CT values observed for each sample type demonstrate the consistent performance of the kit in yielding DNA suitable for PCR amplification. Panel B. Analysis by gel electrophoresis. Agarose gel electrophoresis of PCR products from the 8 different samples of mouse liver genomic DNA indicated that DNA quality and yield were comparable for each.
At-a-glance
| Technology | Silica membrane |
| Format | 96-well plate |
| Processing | Manual or automated, vacuum or centrifugation |
| Starting material | ≤10 mg tissue ≤15 mg paraffin |
| Fragment size | 50 bp–approx. 5 kb |
| Typical yield | Depends on the amount and quality of the sample |
| A260/280 | Strongly depends on sample quality |
| Elution volume | 100 µl |
| Preparation time | ~60 min/plate (excl. lysis) |
| Binding capacity | 20 µg |
Related Products
Genomic DNA purification articles
NucleoSpin Tissue
Reliably obtain high-quality genomic DNA from a wide array of sample types.
NucleoSpin DNA RapidLyse
Speed up your workflow with a unique lysis chemistry that efficiently releases gDNA from cells, tissues, and organs.
NucleoSpin Tissue XS
Get highly concentrated DNA from small inputs, such as biopsy materials, forensic samples, or FACS-sorted cells.
NucleoSnap DNA Plasma
Obtain high-quality cell-free DNA from larger sample volumes.
NucleoSpin Plasma XS
Capture circulating DNA from low inputs of plasma or serum.
NucleoMag DNA Plasma
Isolate cell-free DNA from up to 10 ml of plasma using scalable magnetic-bead technology.
NucleoMag Pathogen
Efficiently isolate viral and bacterial nucleic acids from a wide array of inputs.
NucleoMag Virus
Automate your workflow for high-throughput isolation of viral nucleic acids from serum or plasma.
NucleoMag Blood
Isolate genomic DNA from whole blood using scalable magnetic-bead technology.
NucleoBond HMW DNA
The NucleoBond HMW DNA kit enables isolation of high-quality genomic DNA up to 200 kb in size for long read-sequencing applications.
NucleoMag DNA FFPE
Purify DNA from FFPE tissue samples using paramagnetic beads.
NucleoSpin 96 DNA FFPE
High-throughput purification of DNA from FFPE samples in a 96-well format.
NucleoMag DNA Swab
The NucleoMag DNA Swab kit enables high-throughput isolation of genomic DNA from diverse swab samples.
NucleoMag VET
Efficiently isolate viral and bacterial nucleic acids from common veterinary samples.
NucleoMag 384 Plant
Scale up your plant DNA extraction workflow with parallel processing of 384 samples.
NucleoSpin DNA Plasma Midi
Avoid tedious centrifugation steps and process up to 24 samples in parallel.
Takara Bio USA, Inc.
United States/Canada: +1.800.662.2566 • Asia Pacific: +1.650.919.7300 • Europe: +33.(0)1.3904.6880 • Japan: +81.(0)77.565.6999
FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. © 2023 Takara Bio Inc. All Rights Reserved. All trademarks are the property of Takara Bio Inc. or its affiliate(s) in the U.S. and/or other countries or their respective owners. Certain trademarks may not be registered in all jurisdictions. Additional product, intellectual property, and restricted use information is available at takarabio.com.
