Stool and Soil Genomic DNA Kit

Stool and Soil Genomic DNA Kit provides a simple and fast method to isolate and purify genomic DNA from stool or soil samples. A unique lysis buffer is used to achieve effective removal of inhibitors and contaminants in solid and liquid samples, and magnetic beads are used for specific binding to DNA. The isolated DNA is suitable for PCR, qPCR and Next Generation Sequencing. This kit is compatible with high-throughput automated nucleic acid purification instruments using magnetic rods.

Kit contents:

• Lysis Buffer: 40 ml
• Lysis Enhancer: 2.5 ml
• Defoaming Reagent: 400 µl
• Precipitation Buffer: 12 ml
• Humic Acid Removal Solution: 10 ml
• Binding Buffer: 30 ml
• Cleaning Buffer: 40 ml
• Wash Buffer: 20 ml
• Elution Buffer: 10 ml
• Magnetic Stool and Soil Beads: 900 µl
• Lysis Tubes: 50

Materials required but not provided:

• Magnetic stand
• Sterile pipettes and pipette tips
• 1.5 ml sterile centrifuge tubes
• Centrifuge
• Vortex mixer
• Water bath (optional)

Target	Stool and Soil Genomic DNA Kit
Storage	Store the Humic Acid Removal Solution between 2-8 °C, and all other components at room temperature.
Shelf Life	24 months.
Availability	Shipped within 10-20 working days.
Note	THIS PRODUCT IS FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC, THERAPEUTIC OR COSMETIC PROCEDURES. NOT FOR HUMAN OR ANIMAL CONSUMPTION.
Directions for use	Reagent preparation: To prepare the Working Cleaning Buffer, add 10 ml of 100% ethanol to 40 ml Cleaning Buffer. To prepare the Working Wash Buffer, add 80 ml of 100% ethanol to 20 ml Wash Buffer. To prepare the Working Lysis Reagent, add Lysis Buffer, Lysis Enhancer, and Defoaming Reagent in a 700:40:4 ratio (e.g. add 700 µl Lysis Buffer, 40 µl Lysis Enhancer, and 4 µl Defoaming Reagent to prepare 744 µl of Working Lysis Reagent). Genomic DNA extraction and Isolation: Add sample into a Lysis Tube. For solid samples, add 250-500 mg of soil sample or 100 mg of stool sample. For liquid samples, add 200-400 µl of soil sample or 100-200 µl of stool sample. The sample quantity can be adjusted if the sample water content is low. Add 744 µl of Working Lysis Reagent to the tube. Close the lid. Vortex the tube at maxmimum speed for 10 minutes to thoroughly homogenize the sample. (Optional: incubate in a 70 °C water bath for 5-10 minutes to enhance buffer penetration.) Centrifuge the tube at 12,000 × g for 3 minutes, then transfer 600 µl of supernatant into a new 1.5 ml sterile centrifuge tube. Add 250 µl of Precipitation Buffer and 150 µl of Humic Acid Removal Solution. Mix well by vortexing. Centrifuge at 12,000 × g for 3 minutes. Transfer 500-600 µl of supernatant to a 1.5 ml sterile centrifuge tube, taking care to avoid transferring any precipitate at the bottom. Add 500 µl of Binding Buffer and mix well. Vortex the Magnetic Stool and Soil Beads thoroughly, then add 15 µl of Magnetic Stool and Soil Beads to the tube. Vortex for 5 minutes and then place the tube on a magnetic stand. Gently invert the magnetic stand and rotate the tube so that all the magnetic beads collect on one side of the magnetic stand. Allow to stand until the solution becomes clear. Discard the supernatant carefully, taking care to avoid pipetting the beads. Add 800 µl of Working Cleaning Buffer and vortex for 2 minutes. Place the tube on a magnetic stand until the solution becomes clear. Discard the supernatant carefully, taking care to avoid pipetting the beads. Add 700 µl of Working Wash Buffer and vortex for 2 minutes. Place the tube on a magnetic stand until the solution becomes clear. Repeat once more for a total of 2 times. Discard as much supernatant as possible. Air-dry the uncapped beads at room temperature for 5-10 minutes to evaporate the residual ethanol completely. Add 50-100 µl Elution Buffer. Mix by pipetting up and down. Incubate at 65 °C for 10 minutes, pipetting up and down 2-3 times during this period to mix the solution. Place the tube on a magnetic stand until the solution becomes clear. Transfer the supernatant carefully into a new sterile centrifuge tube. Store the purified gDNA at -20°C. Notes: Avoid repeated freeze/thaw cycles, as this can affect the quality of the extracted DNA. The Magnetic Stool and Soil Beads must be thoroughly vortexed before use. Use sterile nuclease-free tubes and pipette tips to avoid DNase contamination. Before extracting and weighing the sample, vortex to completely homogenize the sample.

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