Description
With the EZ2 PowerFecal Pro DNA/RNA Kit, you can automate the isolation of nucleic acids from stool on the EZ2 Connect. The kit uses second-generation Inhibitor Removal Technology® (IRT), resulting in high yields of highly pure DNA, RNA or total nucleic acids, which can be used immediately in downstream applications, including RT-PCR, qPCR, dPCR and next-generation sequencing. Proven bead beating technology and lysis chemistry complete the user-friendly, proprietary protocol.
EZ2 PowerFecal Pro DNA/RNA Kit enables the automated extraction of high-yield (Figures 1 and 2) and high-purity (Figure 3) DNA, RNA or total nucleic acids from stool samples. The kit efficiently removes PCR inhibitors for highly pure nucleic acids, performing as well as the spin column PowerFecal Pro protocols (Figure 4).
The EZ2 PowerFecal Pro DNA/RNA Kit efficiently isolates DNA, RNA or total nucleic acids from stool samples, depending on the isolation protocol you use. After the combined lysis and inhibitor removal step, you transfer the eluate onto the EZ2 Connect and can choose one of the three extraction protocols: only DNA, only RNA or total nucleic acids. The additional enzymatic digestion steps for DNA-only and RNA-only extractions are performed on the instrument. Respective enzymes, RNase A (17,500 U) (cat.no. 19101) and RNase-free DNase Set (50) (cat.no. 79254) need to be purchased separately.
The recommended amount of starting material is 50 mg of stool with a maximum of 100 mg. Please note that the use of phenol chloroform isoamyl alcohol is crucial for the depletion of the downstream inhibitors and high nucleic acid yields. At the same time, it also deactivates RNase allowing the extraction of high-quality RNA. Sample storage and preservation The yield and integrity of nucleic acids isolated from microbes in stool is greatly influenced by the state of the digestive system, diet of the individual and the length of time between collection of the sample and preservation. To optimize the quality of nucleic acids from stool, process the sample as quickly as possible after collection. To stabilize stool samples at room temperature, use the PowerProtect DNA/RNA reagent (more information). Automated processing on the EZ2 Connect The EZ2 Connect instruments can perform all steps following lysis and disruption of the sample. This automation is based on magnetic particle technology and includes nucleic acid binding, optional enzymatic digestion (when only DNA or RNA are isolated), washing and elution. Up to 24 samples can be processed in a single run. Magnetic-particle technology combines the speed and efficiency of silica-based nucleic acid purification with the convenient handling of magnetic particles. Nucleic acids are isolated from lysates through binding to the silica surface on the particles in the presence of a chaotropic salt. The particles are subsequently separated from the lysates using a magnet. Additional washing steps remove any residual contaminants. Finally, nucleic acids are efficiently eluted. All enzymatic digestions, if isolating DNA or RNA only, are automatically performed during the extraction procedures.
