The NucleoSpin FFPE DNA kit provides a convenient, reliable, and fast method to isolate DNA from formalin-fixed, paraffin-embedded (FFPE) tissue specimens. The procedure omits the use of flammable and malodorous xylene or d-limonene commonly used for deparaffinization. Further, the procedure omits the difficult removal of organic solvent from often barely visible tissue pellets. NucleoSpin FFPE DNA employs the odorless Paraffin Dissolver (patent pending) and allows efficient lysis in a convenient two-phase system.**
First, the paraffin of FFPE sections is dissolved in the Paraffin Dissolver. Tissue is then digested by proteinase to solubilize the fixed tissue and release DNA into solution. Subsequently, heat incubation with a specially designed buffer effectively eliminates crosslinks from the previously released DNA. After addition of ethanol, the lysate is applied to the NucleoSpin FFPE DNA Column. DNA is bound to the silica membrane. Two washing steps help to remove salts, metabolites, and macromolecular cellular components. Pure DNA is finally eluted under low ionic strength conditions in a small volume (5–30 µl) of Elution Buffer BE, yielding highly concentrated DNA.
Isolated DNA is high in concentration, formaldehyde crosslinks are reversed efficiently, and the DNA quality of the sample is well preserved during the complete process (deparaffinization, decrosslink, purification). Thus, isolated DNA shows good performance in typical downstream applications like PCR.
**Users preferring xylene deparaffinization will find protocols utilizing xylene for deparaffinization in the manual. Deparaffinization using xylene shows the same efficiency as deparaffinization using Paraffin Dissolver. Xylene is not included in the kit.