EZ2 AllPrep DNA/RNA FFPE Kit

For automated and simultaneous purification of DNA and RNA from formalin-fixed, paraffin-embedded (FFPE) tissue samples

Features

  • Patented technology to extract DNA and RNA from the same sample – no need to divide the sample
  • Elution of DNA and RNA in separate fractions offers convenient downstream analysis
  • Reduces sample consumption
  • Optimizes crosslink removal for both DNA and RNA
  • No overnight proteinase K incubation
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EZ2 AllPrep DNA/RNA FFPE Kit (48)

Cat. No. / ID: 954734

For 48 preps: EZ2 AllPrep DNA/RNA FFPE cartridge, disposable filter-tips and tip-holders, tubes, Paraffin Removal Solution, Proteinase K, DNase, RNase-free water and buffers
$776.00
The EZ2 AllPrep DNA/RNA FFPE Kit (48) is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Product Details

The EZ2 AllPrep DNA/RNA FFPE Kit is used with the EZ2 Connect instrument. It automates the extraction of both DNA and RNA, from exactly the same piece of sample in two distinct elution fractions.

The kit technology allows the reversal of crosslinking caused by the sample’s exposure to formalin without overnight proteinase K incubation.

Performance

The EZ2 AllPrep DNA/RNA FFPE Kit is designed for automated purification of DNA and RNA from FFPE samples, using magnetic-bead technology and prefilled cartridges.

Principle

DNA and RNA extraction from the same sample is needed to compare genomic and transcriptomic data reliably. Usually, this is done by splitting the sample into two separate reactions/tubes/containers. The extractions are then processed separately. Thus, DNA is extracted from one piece of the sample, while RNA is extracted from the other piece.

Conventional methods have two major disadvantages: First, the amount of nucleic acids that can be extracted from the sample is cut into half. Second, because DNA is extracted from one group of cells, and RNA is extracted from a separate group, there can be differences in the properties of the cell sources.

The patented AllPrep DNA/RNA FFPE procedure incubates FFPE tissue samples in a lysis buffer that releases RNA into the supernatant and leaves DNA in the pellet. The RNA and DNA are then purified in separate tubes but within the same automated run using one cartridge in the EZ2 Connect instrument. This way, DNA and RNA can be isolated from as many as 24 samples, all at the same time.

Crosslink removal

Extended crosslink removal at high temperature reverses more crosslinks, but it also increases nucleic acid fragmentation. RNA is more susceptible to heat-induced fragmentation than DNA – but it is also less affected by crosslinks.

Therefore, the optimal conditions for decrosslinking DNA and RNA are different.

By generating separate lysates for RNA and DNA, the EZ2 AllPrep DNA/RNA FFPE workflow allows optimal crosslink removal conditions for each nucleic acid type.

Procedure

EZ2 AllPrep DNA/RNA FFPE technology solubilizes and incubates a single FFPE sample to release RNA into the supernatant while DNA is left in the pellet. The supernatant is incubated to remove RNA crosslinking. Finally, the tubes that separately contain the supernatant and pellet(s) are transferred to EZ2 Connect.

The EZ2 Connect automates all remaining steps of the workflow. These include an additional lysis and crosslink-removal step for DNA, binding of nucleic acids to magnetic particles, removal of contaminants with multiple washing steps, and elution of DNA and RNA into different tubes (see  EZ2 AllPrep DNA/RNA FFPE workflow).

See figures

Applications

DNA and RNA purified with the EZ2 AllPrep DNA/RNA FFPE Kit can be used for real-time PCR, Pyrosequencing and other downstream applications.

Supporting data and figures

Specifications

FeaturesSpecifications
Applications(RT-)PCR, (RT-)qPCR, NGS, genotyping, microarray analysis
Elution volumeRNA: 50–60 µl; DNA: 100–120 µl
Purification of total RNA, miRNA, poly A+ mRNA, DNA or proteinRNA and DNA
TechnologyMagnetic particle technology
Number of preps per run1–24 samples per run
Main sample typeFFPE tissue samples
Sample amountmax. 4*10 µm sections or 2*20 µm sections
YieldVaries depending on sample type, as well as on storage and fixation conditions

Resources

Kit Handbooks (1)
Quick-Start Protocols (1)