QIAamp DNA FFPE Tissue Kit
For purification of genomic DNA from formalin-fixed paraffin-embedded tissues
- Rapid purification of high-quality, ready-to-use DNA
- Consistent, high yields
- Complete removal of contaminants and inhibitors
Cat. No. / ID: 56404
The QIAamp DNA FFPE Tissue Kit is specially designed for purifying DNA from formalin-fixed, paraffin-embedded tissue sections. The kit uses special lysis conditions to release DNA from tissue sections and to overcome inhibitory effects caused by formalin crosslinking of nucleic acids. The kit uses QIAamp MinElute spin columns for purification of high-quality DNA in small volumes. Purification of DNA using the QIAamp DNA FFPE Tissue Kit can be automated on the QIAcube Connect.
This kit will be discontinued soon. Try the next-generation QIAamp DNA FFPE Advanced Kits for efficient recovery from DNA and optional removal of artefacts.
The QIAamp DNA FFPE Tissue Kit uses well-established QIAamp MinElute technology for purification of genomic and mitochondrial DNA from small sample volumes or sizes. The kit combines the selective binding properties of a silica-based membrane with flexible elution volumes of between 20 and 100 μl.
Specially optimized lysis conditions allow genomic DNA to be efficiently purified from FFPE tissue sections without the need for overnight incubation. Incubation at an elevated temperature after proteinase K digestion partially removes formalin crosslinking of the released DNA, improving yield as well as DNA performance in downstream assays. Note that DNA isolated from FFPE samples is usually of lower molecular weight than DNA from fresh or frozen samples. The degree of fragmentation depends on the type and age of the sample and the conditions used for fixation.
The QIAamp DNA FFPE Tissue procedure consists of 6 steps: remove paraffin, lyse, heat, bind, wash, and elute (see flowchart " Procedure"). After sample lysis, the simple QIAamp DNA FFPE Tissue procedure, which is highly suited for simultaneous processing of multiple samples, yields pure DNA in less than 30 minutes.
The QIAamp DNA FFPE Tissue Kit is specially designed for purifying DNA from formalin-fixed paraffin-embedded tissue.
Genomic DNA was purified from different FFPE tissue samples using the QIAamp DNA FFPE Tissue Kit. Products of the prnp gene (78 bp, 464 bp, and 727 bp) were amplified by real-time quantitative PCR.
|Applications||Real-time PCR, STR analysis, LMD-PCR|
|Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein||Genomic DNA, mitochondrial DNA|
|Elution volume||20-100 µl|
|Main sample type||Formalin fixed paraffin embedded tissue samples|
|Sample amount||Up to 8 sections, each with a thickness of up to 10 µm and a surface area of up to 250 mm2|
The MinElute spin columns included in the following kits should be stored at 2–8°C upon arrival: AllPrep DNA/RNA Micro, EpiTect Fast DNA Bisulfite, EpiTect Fast FFPE Bisulfite, EpiTect Fast LyseAll Bisulfite, EpiTect Plus DNA Bisulfite, EpiTect Plus FFPE Bisulfite, EpiTect Plus LyseAll Bisulfite, exoRNeasy Serum/plasma Maxi, exoRNeasy Serum/Plasma Midi, GeneRead DNA FFPE, GeneRead rRNA Depletion, GeneRead Size Selection, MinElute Gel Extraction, MinElute PCR Purification, MinElute Reaction Cleanup, miRNeasy FFPE, miRNeasy Micro, miRNeasy Serum/Plasma, QIAamp DNA FFPE, QIAamp DNA Investigator, QIAamp DNA Micro, QIAamp MinElute Media, QIAamp MinElute Virus Spin, QIAamp MinElute Virus Vacuum, RNeasy FFPE, RNeasy Micro, RNeasy Plus Micro.
Short-term storage (up to 4 weeks) at room temperature (15–25°C) does not affect the performance. However, for optimal performance and quality, storage temperature should not exceed 25°C.
The composition of Buffer ATE is:
- 10 mM Tris-Cl pH 8.3
- 0.1 mM EDTA
- 0.04% NaN3 (sodium-azide)
It is strongly advised to follow the recommendations for preparing sample material provided in the corresponding Protocol Sheet to ensure reliable results.
Plasma: It is recommended to perform plasma separation immediately after blood collection when using EDTA or citrate as anticoagulant to prevent the release of genomic DNA into the plasma fraction.
Urine: Because circulating cell-free DNA in non-stabilized urine samples is rapidly degraded after sample collection due to high nuclease activity, eluates may contain no DNA or exhibit low DNA concentration. Therefore, it is recommended to stabilize urine samples. Even when using stabilized urine, it is recommended to perform a centrifugation step immediately after stabilization to prevent the release of genomic DNA from cells. Alternatively, non-stabilized urine samples can be processed immediately after collection and centrifugation using ATL-pretreatment and automated DNA extraction as described in the corresponding Protocol Sheet.
Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.
Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).