QIAamp DNA FFPE Tissue Kit

For purification of genomic DNA from formalin-fixed paraffin-embedded tissues

Features

Rapid purification of high-quality, ready-to-use DNA
Consistent, high yields
Complete removal of contaminants and inhibitors

Product Details

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.

Try the next-generation QIAamp DNA FFPE Advanced Kits for efficient recovery from DNA and optional removal of artefacts.

Performance

The QIAamp DNA FFPE Tissue Kit purifies DNA that can be used in a wide range of downstream applications, such as single nucleotide polymorphism (SNP) and short-tandem repeat (STR) genotyping and pharmacogenomic research. DNA purified with the QIAamp DNA FFPE Tissue Kit can also be used in PCR (see figure "").

See figures

PCR analysis of DNA purified from FFPE tissue samples.

Principle

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.

Procedure

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.

See figures

QIAamp DNA FFPE Tissue Kit procedure.

Applications

The QIAamp DNA FFPE Tissue Kit is specially designed for purifying DNA from formalin-fixed paraffin-embedded tissue.

Supporting data and figures

PCR analysis of DNA purified from FFPE tissue samples.

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.

Specifications

Features	Specifications
Applications	Real-time PCR, STR analysis, LMD-PCR
Processing	Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein	Genomic DNA, mitochondrial DNA
Elution volume	20-100 µl
Format	Spin column
Technology	Silica technology
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

Resources

Publications

Gender-related survival differences associated with EGFR polymorphisms in metastatic colon cancer.

Press OA; Zhang W; Gordon MA; Yang D; Lurje G; Iqbal S; El-Khoueiry A; Lenz HJ;

Cancer Res; 2008; 68 (8):3037-42 2008 Apr 15 PMID:18413774

Discrepancy between CYP2D6 phenotype and genotype derived from post-mortem dextromethorphan blood level.

Bailey B; Daneman R; Daneman N; Mayer JM; Koren G;

Forensic Sci Int; 2000; 110 (1):61-70 2000 May 8 PMID:10802201

FAQ

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.

FAQ ID - 3560

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.).

FAQ ID -728