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. To achieve effective removal of inhibitors and contaminants in solid and liquid samples, unique lysis buffer is designed 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 A: 50 ml
• Lysis Buffer B: 30 ml
• Precipitation Buffer: 12.5 ml
• Binding Buffer: 10 ml
• Cleaning Buffer: 10 ml
• Wash Buffer: 12 ml
• Elution Buffer: 10 ml
• Magnetic Stool and Soil Beads: 2 ml
• Glass Beads: 12.5 g

Target	Stool and Soil Genomic DNA Kit
Storage	Store the Magnetic Stool and Soil Beads between 2-8 °C for up to one year, and all other components at room temperature (15-25 °C) for up to one year.
Availability	Shipped within 10-20 working days.
Note	This product is for research use only.
Directions for use	Reagent preparation: To prepare the Working Binding Buffer, add 30 ml of 100% isopropanol to 10 ml Binding Buffer. If a precipitate has formed in the Binding Buffer, incubate at 37 °C before use. To prepare the Working Cleaning Buffer, add 10 ml of 100% ethanol to 10 ml Binding Buffer. To prepare the Working Wash Buffer, add 48 ml of 100% ethanol to 12 ml Wash Buffer. Prepare a water bath (or other heating apparatus) set to 70 °C. Stool genomic DNA extraction: Pretreatment: Add 100-300 mg of solid sample or 100-200 µl of liquid sample into a 1.5 ml centrifuge tube. Add 0.25 g glass beads and 1 ml of Lysis Buffer A. Vortex the tube until the stool sample is thoroughly homogenized. (Optional: if RNA-free genomic DNA is required, add 20 µl of RNase A (abx098139). Allow to stand for 3 minutes at room temperature). Incubate at 70 °C for 10 minutes. Centrifuge the tube at 12,000 × g for 2 minutes, then transfer the supernatant into a new 1.5 ml sterile centrifuge tube. Lysis: Add 250 µl of Precipitation Buffer and mix well by vortexing. Allow to stand on ice for 5 minutes. Centrifuge at 12,000 × g for 2 minutes, and transfer up to 600 µl of supernatant to a 1.5 ml sterile centrifuge tube. If turbidity is still observed in the supernatant, centrifuge at 12,000 × g for 1 minute and transfer the supernatant to a 1.5 ml sterile centrifuge tube. Follow the steps in the Isolation procedure below. Soil genomic DNA extraction: Pretreatment: Weigh 250 mg soil sample in a 1.5 ml centrifuge tube. Add 0.25 g glass beads and 600 µl of Lysis Buffer B. Vortex for 15 minutes. Centrifuge the tube at 12,000 × g for 2 minutes, then transfer the supernatant into a new 1.5 ml sterile centrifuge tube. Lysis: Centrifuge the tube at 12,000 × g for 2 minutes, then transfer the supernatant to a new 1.5 ml sterile centrifuge tube. Add 200 µl of Precipitation Buffer and mix well by vortexing. Allow to stand on ice for 5 minutes. Centrifuge at 12,000 × g for 2 minutes, and transfer up to 600 µl of supernatant to a 1.5 ml sterile centrifuge tube. Follow the steps in the Isolation procedure below. Isolation procedure: Add 500 µl of Working Binding Buffer and 30 µl of Magnetic Stool and Soil Beads to the tube. Vortex for 1 minute and then allow to stand at room temperature for 2 minutes. Repeat 3 times for a total of 4 times. Place the tube on a magnetic stand for 5 minutes, until the solution becomes clear. Discard the supernatant carefully, taking care to avoid pipetting the beads. Add 400 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 600 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. Pipette supernatant as thoroughly 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 and vortex for 30 seconds. Incubate at 65 °C for 10 minutes, vortexing 2-3 times during this period. 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: Carry out all magnetic separation steps at room temperature. Beads must be vortexed before use. Use sterile tubes and pipette tips to avoid DNase contamination. Stool samples in ethanol should be centrifuged and washed with sterilized distilled water 2-3 times to remove the ethanol before use. Before extracting, vortex to completely homogenize the sample. To help obtain high yield, samples should be kept under optimal conditions. To help obtain high extraction purity, avoid pipetting the precipitate when transferring the supernatant.

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