Rapid, High-Throughput Isolation of PCR-Ready Genomic DNA from Soil Samples The FastDNA™-96 Soil Microbe DNA Kit is designed for the simple and rapid isolation of humic-free, PCR-quality DNA from microbes in soil in approximately 50 minutes. Gram(+/-) bacteria, fungi, plant and animal tissues, algae, spores and many other members of a soil population including clay, sandy, silty, peaty, chalky, and loamy soils, are efficiently lysed in approximately 60 seconds with the FastPrep-96™ Instrument. Up to 130 mg of soil 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 all soil organisms including historically difficult sources such as eubacterial spores and endospores, gram (+/-) bacteria, yeast, algae, nematodes, and fungi, 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 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.
Matrix Y, 0.5 mm diameter Yttria-Stabilized Zirconium Oxide
Molecule to be Enriched
DNA
Kit Components
FastDNA-96™ Lysing Matrix Rack (0.5 mm Beads) 2 x 96-well rack, Lysis Buffer 2 x 40 ml, Soil DNA Binding Solution 150 ml, Binding Plate Pre-Wash Buffer 50 ml, Soil DNA Wash Buffer 100 ml, Elution Solution 2 x 10 ml, Inhibitor Plate Prep Solution 30 ml, Deep-Well Plate 2 each, MP-96 Binding Plate 2 each, MP-96 Inhibitor Removal Plate 2 each, Collection Plate 2 each, Elution Plate 6 each, Foil Plate Cover 4 each, User manual 1 each, MSDS 1 each, Certificate of Analysis 1 each
Materials Required but not Supplied
FastPrep-96™ Instrument or alternative high speed disruptor, Centrifuge with a swing-bucket style rotor that can spin up to 3,500 – 5,000 rpm, Swing-bucket centrifuge microplate adaptors for 96-well plates (pair), Plate shaker, Sterile water, beta-mercaptoethanol
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