The FastDNA-96™ Fecal DNA Kit isolates bacterial, protist, or host DNA from up to 80 mg of wet or dry stool, and even from caecum samples, through a purification process that eliminates PCR inhibitors. Fecal microbes, fungi, parasites, and other stool organisms are first lysed in approximately 60 seconds with the FastPrep-96™ Instrument.
Up to 80 mg of wet or dry feces are placed into each 1.2 mL tube of a FastDNA-96™ Lysing Matrix Rack (96-deep well plate) containing 0.5 mm specially stabilized Zirconium oxide lysing particles, which are designed to efficiently lyse a wide variety of eukaryotic and prokaryotic organisms present in mammalian stool (i.e. humans, primates, birds, rodents, cattle, canine, feline, porcine, etc.), while in the presence of a specially formulated lysis solution. The cleared lysate is then applied to a binding matrix to isolate the DNA, and spin column plates are used to simplify liquid handling. Eluted DNA is then applied to the MP-96 Inhibitor Removal Plate which removes humic acids, polyphenols that can inhibit PCR. Purified DNA is eluted in an EDTA-free, DNA elution solution, and is ready for downstream applications including digestion, electrophoresis, methylation and PCR (A260/A280 ratios = 1.8). Yield is typically 5 µg of total DNA eluted in 50-100 µL of elution solution. This procedure can be performed in minutes, and there is no need for organic denaturants or proteinases.
The FastDNA-96™ Fecal DNA Kit quickly and efficiently isolates inhibitor-free, PCR-ready genomic DNA from stool samples (i.e. humans, primates, birds, rodents, cattle, canine, feline, porcine, etc.).