EZ1&2 DNA FFPE UNG Kit

QIAGEN offers this kit for the automated purification of genomic DNA from formalin-fixed, paraffin-embedded (FFPE) tissues for molecular biology applications. The kit is intended to be used on EZ1 Advanced XL or EZ2 Connect instruments, including EZ2 Connect, EZ2 Connect Fx, and EZ2 Connect MDx. The procedure removes paraffin without the use of hazardous solvents and without the need to trim excess paraffin from the FFPE block in advance.

Two Proteinase K digestion steps—one before and one after DNA de-crosslinking—ensure complete lysis of difficult-to-lyse tissue and facilitate the recovery of high amounts of amplifiable DNA. Following the initial Proteinase K digestion and elevated-temperature incubation to remove cross-links, the reaction mixture is diluted to permit removal of deaminated cytosine residues using Uracil-N-Glycosylase (UNG). This UNG step addresses the most common artifact in FFPE tissues, where cytosine deaminates to uracil and produces C→T conversion artifacts during amplification, which can lead to false results in sensitive methods such as next-generation sequencing or digital PCR with limited starting material. Prior to DNA binding, a second Proteinase K digestion improves lysis efficiency, particularly for difficult-to-lyse samples. DNA binds to magnetic particles, contaminants are washed away, and DNA is eluted in Buffer AVE. Isolated DNA is compatible with PCR, digital PCR, and NGS workflows, and may be stored long term at −30 to −15°C. Up to 24 samples may be processed in a single run.

The kit is shipped at ambient temperature; buffers and reagents are stable at room temperature (15–25°C), and reagent cartridges should not be frozen. The UNG component is shipped on dry ice and should be stored at −30 to −15°C upon receipt. Sample material consists of 5 to 10 µm sections cut from an FFPE block, with the starting material calculated from tissue surface area, number of sections, and section thickness. Recommended practice includes fixation in 4%–10% formalin, keeping fixation time to a minimum, and thoroughly dehydrating samples after fixation to limit nucleic acid fragmentation.