The NZY Stool RNA Isolation kit is engineered to ensure efficient extraction of RNA, including smaller RNA species, from both fresh and frozen stool samples. Utilizing the combined action of Buffer NST NZYol and a specialized Lysis Buffer NSTL, the kit offers an effective chemical disruption method for the stool sample and the microbes contained therein. This chemical disruption is supplemented by mechanical lysis using NZYSpin Stool Bead Tubes (equipped with ceramic beads) and a 10-minute shaking on a Vortex. Importantly, the homogenization of the sample material does not necessitate any enzymatic reactions such as protease digestion. The kit’s procedure involves separating the undissolved sample material and ceramic beads through brief centrifugation. This is followed by adding Buffer NSTB (binding buffer) to further clear the lysate. The clarified supernatant is subsequently passed through the NZYSpin RNA Stool Filter Columns (red rings), effectively eliminating elements from stool samples that could hinder RT-PCR processes. After adjusting the binding conditions by introducing additional Buffer NSTB to the NZYSpin RNA Stool Filter Columns flow-through, the sample is loaded onto NZYSpin RNA Stool Columns (light blue rings). Any lingering contaminants, including complex polysaccharides, bile salts, and other PCR inhibitors, are then effectively removed through a thorough washing procedure utilizing Buffer NSTB along with Wash Buffers NSTW1, NSTW2, and NSTW3. Residual DNA is eliminated during the washing stages via an on-column DNA digestion process involving rDNase. Following a drying phase, the RNA, now ready for use, can be eluted using RNase-free water.