產(chǎn)地：Qiagen
貨號：56404
Principle and procedure
The QIAamp DNA FFPE Tissue procedure consists of 6 steps （see flowchart "QIAamp DNA FFPE Tissue Procedure"）:
Remove paraffin: paraffin is dissolved in xylene and removed
Lyse: sample is lysed under denaturing conditions with a short proteinase K digestion
Heat: incubation at 90°C reverses formalin crosslinking
Bind: DNA binds to the membrane and contaminants flow through
Wash: residual contaminants are washed away
Elute: pure, concentrated DNA is eluted from the membrane
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.
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.
DNA is eluted in Buffer ATE or water and is immediay ready for use in amplification reactions or for storage at –20°C. Purified DNA is free of proteins, nucleases, and other impurities.