The Soil DNA Kit was created specifically for isolating genomic DNA from soil samples and other environmental samples. Its quick and easy workflow allows for DNA isolation in under half an hour and removes humic substances and DNA PCR inhibitors. The Kit offers high purity and stable quality DNA free of proteins, humic substances, and color from compost, manure, and sediment thanks to its unique silica-membrane technology and specific buffer system. Purified DNA can be employed directly in molecular biological experiments like PCR, restriction enzyme digestion, and the creation of DNA libraries. All soil specimen or other environmental specimen is acceptable for the species segment of the specimens.
Storage:
Store at room temperature (15-25℃)
Components:
Specifications:
| Features | Fast and convenient workflow enables DNA isolation within 30min. Eliminates humic substances and PCR inhibitors for DNA. |
| Application | The purified DNA can be used directly for molecular biological experiments such as PCR, restriction enzyme digestion and DNA library construction. |
| Species Category | all soil samples or other environmental samples |
Ensure that Buffer PWS have been prepared with appropriate volume of ethanol as indicated on the bottle tag and shake thoroughly.
- Add 750 μl Buffer SA and 0.25 g Glass beads to a 2ml microcentrifuge tube.
- Add 0.25 g soil sample to the 2 ml microcentrifuge tube, mix by vortex for 15 sec.
- Add 60 μl Buffer SC to the sample and mix by vortex for 10 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.
- Add 250 μl Buffer HA, mix by vortex for 5 sec, and incubate the tube at 4 °C for 5 min.
- Centrifuge at 12,000 rpm (~13,400 × g) for 1 min, transfer the supernatant to a new 2 ml microcentrifuge tube, add 200 μl Buffer HB and mix, incubate the tube at 4 °C for 5 min.
- Centrifuge at 12,000 rpm (~13,400 × g) for 1 min, transfer the supernatant to a new 2 ml microcentrifuge tube, add 1,200 μl Buffer GF and mix by inverting.
Note: Avoid pipetting precipitate when transferring supernatant, or else the purity of product would be affected. - Transfer 700 μl solution from step 7 to a Spin Column CB3 (place the column in a collection tube), centrifuge at 12,000 rpm (~13,400 × g) for 30 sec, discard the flow-through, put the column back to the collection tube.
Note: The capacity of Spin Column CB3 is 700 μl, if the sample volume exceeds 700 ul, centrifuge successive aliquots in the same column. Discard the flow -through after each centrifugation. - Add 500 μl Buffer PWS ( Ensure that ethanol (96-100%) has been added ) to the Spin Column CB3, centrifuge at 12,000 rpm (~13,400 × g) for 30 sec, discard the flow -through, put the column back to the collection tube.
- Add 500 μl 70% ethanol to the Spin Column CB3, centrifuge at 12,000 rpm (~13,400 × g) for 30 sec, discard the flow -through, put the column back to the collection tube.
- Centrifuge at 12,000 rpm (~13,400 × g) for 2 min, discard the flow-through.
- Incubate the Spin Column CB3 at room temperature (15 -25°C) for several minutes to completely dry the residual washing buffer in the column. Place the Spin Column CB3 in a new clean microcentrifuge tube, and pipet 50-100 μl Buffer TE directly to the center of the membrane. Incubate at room temperature (15-25°C) for 2-5 min, and then centrifuge for 2 min at 12,000 rpm (~13,400 × g), collect the flow-through to the microcentrifuge tube.
