The Magnetic Soil/Stool DNA Kit was created specifically for isolating genomic DNA from stool specimens, all soil specimens, and other difficult environmental specimens with a high humic acid content, such as compost, sediment, and manure. DNA connects to the silica surface of the magnetic particles, free of proteins and other contaminants, using magnetic-particle technology and a specific buffer solution. Purified DNA can be instantly used in molecular biological experimentations such as restriction analysis, PCR analysis, and next-generation sequencing (NGS). The Kit can be kept at room temperature (15-25°C) and requires approximately 250-500 mg of sample. Stool samples, all soil samples, and other difficult environmental samples with a high humic acid content are all acceptable sample types.
Storage:
Store at room temperature (15-25℃)
Components:
Specifications:
| Sample amount | 250-500 mg |
| Features | Purified genomic DNA can be extracted from stool samples, all soil samples and other difficult environmental samples containing a high humic acid content, such as compost, sediment, and manure. Purified DNA can be used directly for PCR, restriction digestion and NGS. |
| Application | The purified DNA can be used directly for molecular biological experiments such as PCR, restriction digestion and NGS. |
| Sample type | Stool samples, all soil samples and other difficult environmental samples containing a high humic acid content, such as compost, sediment, and manure. |
Ensure that ethanol (96-100%) has been added into Buffer PWD and Buffer RD, isopropanol has been added into Buffer GFA as indicated on the tag of the bottle before use.
- Sample preparation:
1) For soil samples: Transfer 250-500 mg sample to a 2ml microcentrifuge tube (not provided). Add 500μl Buffer SA, 100μl Buffer SC and 0.25g 1 mm Grinding Beads, mix by vortex for 15 min. Centrifuge at 12,000 rpm (~13,400 × g) for 1 min, transfer the supernatant (around 500 μl ) to a new 2 ml microcentrifuge tube.
2) For stool sample: Weigh 250-500 mg stool in a 2 ml microcentrifuge tube (not provided). Add 500μl Buffer SA, 100μl Buffer SC and 0.25g 1 mm Grinding Beads, (there may be high level of residual RNA in stool samples, if necessary, add 10μl RNase A (10 mg/ml, Cat.no. NOR002)) mix by vortex. Centrifuge at 12,000 rpm (~13,400 × g) for 1 min, transfer the supernatant (around 500 μl ) to a new 2 ml microcentrifuge tube. - Add 200μl Buffer SH, mix by vortex for 5 s. Incubate at 4°C for 10 min.
- Centrifuge at 12,000 rpm (~13,400 × g) for 3 min, transfer the supernatant to a new 2 ml microcentrifuge tube. Add 500μl Buffer GFA (Ensure isopropanol has been added), and mix by pipetting or vortex.
- Add 10μl MagAttract Suspension G, mix by pipetting or vortex for 5min.
- Place the centrifuge tube on a magnetic separation device for 30 s. Allow the magnetic particles to completely clear from the solution and then discard the supernatant carefully.
- Remove the centrifuge tube from the magnetic separation device. Add 700 μl Buffer RD (ensure that ethanol (96-100%) has been added), and mix by pipetting or vortex for 5min.
- Place the centrifuge tube on a magnetic separation device for 30 s. Allow the magnetic particles to completely clear from the solution and then discard the supernatant carefully.
- Remove the centrifuge tube from the magnetic separation device. Add 700 μl Buffer PWD (ensure that ethanol (96-100%) has been added), and mix by pipetting or vortex for 3min.
- Place the centrifuge tube on a magnetic separation device for 30 s. Allow the magnetic particles to completely clear from the solution and then discard the supernatant carefully.
- Repeat step 8 and 9.
- Place the centrifuge tube on the magnetic separation device for 5-10 min.
Note: Residual ethanol may inhibit subsequent enzymatic reactions, please ensure the residual ethanol is volatile absolutely. However, overdrying should be avoided, since over-dried DNA is difficult to dissolve. - Remove the centrifuge tube from magnetic separation device. Add 50-100 μl Buffer TB to elute nucleic acids. Mix by pipetting or vortex and incubate at 56°C for 5 min, vortex every 3 min.
- Place the centrifuge tube on the magnetic separation device for 2 min until all the magnetic particles are cleared from the solution. Transfer the supernatant containing purified nucleic acids to a new centrifuge tube and stores it.
