Qiagen, 9026946, EZ1 Advanced XL DNA FFPE Card

Preprogrammed card for purification of DNA using the EZ1 Advanced XL

Features

- High recovery of amplifiable DNA
- Paraffin removal without xylene or similar solvents
- Option to remove uracil (deaminated cytosine) artifacts
- Ready-to-use DNA for PCR, digital PCR (dPCR) and next-generation sequencing (NGS)
- Automated heating and reagent-pipetting pretreatment steps, available on the EZ2 Connect

Product Details

Purify large amounts of amplifiable DNA from hard-to-lyse formalin-fixed, paraffin-embedded (FFPE) tissue. The EZ1&2 DNA FFPE protocol uses double lysis to recover DNA effectively, while the optional uracil-N-glycosylase (UNG) step removes deaminated cytosine artifacts to limit the risk of nucleotide read errors.

The EZ1&2 DNA FFPE protocols are automatable on either the EZ1 Advanced XL or the EZ2 Connect .

The EZ2 Connect features cardless protocols, expanded applications and remote connectivity options. Click here to learn more about the EZ2 Connect , or contact your sales representative to discover how easily you can get an EZ2 Connect for your lab.

Performance

Under UV-vis, fluorometric, and qPCR analysis, most of the samples processed with the EZ1&2 DNA FFPE Kit showed a higher abundance of dsDNA (see figure “ Optimized for PCR performance ”).

For NGS analysis, the EZ1&2 DNA FFPE UNG Kit significantly reduces artificial C→T/G→A transitions, which commonly occur in FFPE material due to cytosine deamination (see “ Artificial C→T/G→A transitions ”). This helps prevent false-positive reports of single-nucleotide variants (SNVs) in NGS (see figure “ Reduction in artifactual C→T | G→A transitions ”).

Principle

The EZ1&2 DNA FFPE Kit maximizes DNA yields from limited sample inputs in two ways: By implementing a two-step lysis procedure, it helps ensure high DNA extraction even from difficult-to-lyse samples. In addition, its wash-free no-solvent deparaffinization technique minimizes the risk of losing of any sample material, which is difficult to avoid with multiple-wash solvent-based deparaffinization methods.

Crosslink removal further increases the amount of recovered amplifiable DNA (see figure “ Optimized for PCR performance ”). In addition, it also makes the recovered DNA especially suitable for NGS analysis (see figure “ Reliable dPCR and NGS results ”).

Procedure

The EZ1&2 DNA FFPE Kit fully automates the bind, wash and elute steps on the EZ1 Advanced XL or the EZ2 Connect. The preparation steps (i.e., Proteinase K digestions, crosslink removal and RNase A digestion) are done manually on the EZ1 Advanced XL; on the EZ2 Connect, many of these preparation steps can be automated (see figure “ EZ1&2 DNA FFPE workflow ”).

Applications

DNA isolated with the EZ1&2 DNA FFPE Kit can be used immediately for PCR, dPCR or NGS. Alternatively, it can be stored at −30 to −15°C.