Maxwell® HT DNA FFPE Isolation System
- Overview
- Protocols
- Specifications
- Resources
High-Throughput Extraction of DNA from FFPE Samples
- Robust, precipitation-free protocol, no chance of lost pellets
- High yields of pure DNA from FFPE samples without using xylene or other hazardous chemicals
- Ideal for use in downstream applications including qPCR and next-generation sequencing (NGS)
Enables rapid extraction of FFPE DNA on liquid handlers
The Maxwell® HT DNA FFPE Isolation System provides a simple and reliable method for high-throughput, rapid isolation of genomic DNA from FFPE (formalin-fixed, paraffin-embedded) tissue samples. The purified DNA can be used directly in a variety of downstream applications, including PCR and next-generation sequencing.
The Maxwell® HT DNA FFPE Isolation System purifies nucleic acid using paramagnetic particles, which provide a mobile solid phase to optimize sample capture, washing and purification of gDNA. The use of paramagnetic particles for DNA capture eliminates the need for centrifugation or vacuum manifolds, making the system suitable for full automation. In addition, the system does not require an organic solvent, making it safe and convenient.
FFPE System Performance Comparison
To demonstrate the performance of this system, Maxwell® HT DNA FFPE purification chemistry was automated on the Tecan Freedom EVO® and ThermoFisher KingFisher™ Flex instruments. Qiagen QIAamp® and GeneRead kits were run using a standard method on a QIAcube instrument. Consecutive FFPE sections of human lung tumor and normal adjacent tissue (NAT) were interspersed among all methods and replicates to minimize sample-based variability. Eluates were quantitated using fluorometric detection and quality was assessed using the ProNex® DNA QC Assay.
Yield of DNA Extracted from FFPE Tissues. Extracted DNA was quantitated using the QuantiFluor® ONE dsDNA System to ensure high accuracy. For both tumor and normal tissue, Maxwell® HT DNA FFPE chemistry yielded consistently greater amounts of DNA than other systems. 
Amplifiability of fragments of various sizes using qPCR. After extraction, the ProNex® DNA QC Assay was used to assess the amplifiability of the DNA using three particular targets as an indication of the DNA integrity and thus suitability for downstream applications, including qPCR and next-generation sequencing. In both sample types and at all amplicon lengths, eluates derived from the Maxwell® HT DNA FFPE Isolation System were shown to provide greater amplification.
| Tissue | Instrument | Uniformity of Coverage (%) | Mean Amplicon Coverage (X) | Reads (millions) |
|---|---|---|---|---|
| Tumor | Tecan | 100.0 | 4818 | 1.08 |
| KingFisher | 99.19 | 6036 | 1.28 | |
| Normal Adjacent | Tecan | 100.0 | 5202 | 1.18 |
| KingFisher | 100.0 | 4074 | 1.05 |
Next-Generation Sequencing Performance of DNA Purified with Maxwell® HT Chemistry. DNA purified using Maxwell® HT chemistry on both Tecan and KingFisher instruments was sequenced using an Illumina MiSeq after enrichment using the Ion AmpliSeq™ Cancer Hotspot Panel v2. Coverage, uniformity and depth were extremely high for both methods and for both tissue types, showing the suitability of these eluates in NGS applications. Furthermore, subsequent variant analysis revealed that low-frequency variants in the KRAS and EGFR genes were detected in the tumor tissue samples, but not in the normal tissue samples.
- Protocols Content
Protocols
Complete Protocol
Maxwell HT DNA FFPE Isolation System Technical Manual
PDF (315 KB)
- Specifications Content
Specifications
What's in the box?
| Item | Part # | Size | Concentration | Available Separately |
|---|---|---|---|---|
|
FFPE Resin |
A220G | 1 × 33ml | ||
|
Mineral Oil |
A244C | 1 × 140ml | ||
|
Lysis Buffer (LBA) |
A287A | 1 × 110ml | ||
|
Buffer B (BWB) |
A288B | 2 × 58.8ml | ||
|
Proteinase K (PK) Solution |
A505E | 1 × 30ml | ||
|
Buffer A (BWA) |
A6371 | 1 × 125ml | View Product | |
|
RNase A Solution |
A797F | 1 × 24ml | 4mg/ml | |
|
Nuclease-Free Water |
P1195 | 1 × 150ml | View Product |
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
- Resources Content
Resources
