The Stool DNA Kit was created specifically for isolating genomic DNA from stool samples in a quick and easy procedure that can be completed in under an hour. The Kit offers high purity and stable quality DNA free of proteins and other organic compounds thanks to its unique silica membrane innovation and particular buffer solution. Purified DNA can be used directly in molecular biological experiments like PCR, Southern hybridization, and the construction of DNA libraries. The sample size required is approximately 180-220 mg.
Storage:
Store at room temperature (15-25℃)
Components:
Specifications:
| Sample amount | 180-220 mg |
| Features | Fast and convenient workflow enables DNA isolation within 1h. |
| Application | The purified DNA can be used directly for molecular biological experiments such as PCR, Southern hybridization and DNA library construction. |
| Sample type | stool samples |
Ensure that Buffer GD and Buffer PW have been prepared with appropriate volume of ethanol (96-100%) as indicated on the bottle and shake thoroughly.
- Weigh 180-220 mg stool in a 2 ml microcentrifuge tube (not provided) and place the tube on ice.
- Add 1.4 ml Buffer GSL, vortex continuously for 1 min until the stool sample is thoroughly homogenized.
- Heat the suspension for 5 min at 70°C.
- Vortex for 15 s, centrifuge at 12,000 rpm (~13,400 × g) for 1 min, transfer 1.2 ml supernatant into a new 2 ml centrifuge tube.
- Add one InhibitEX Tablet to each sample and vortex until the tablet is completely suspended. Incubate the suspension for 1 min at room temperature to allow inhibitors to be adsorbed to the InhibitEX matrix.
- Centrifuge at 12,000 rpm (~13,400 × g) for 3 min.
- Transfer supernatant from step 6 into a new 1.5 ml microcentrifuge tube. Repeat step 6.
- Transfer 200 μl supernatant into a new 1.5 ml microcentrifuge tube. Add 15 μl proteinase K.
- Add 200 μl Buffer GB, and vortex for 15 s.
- Incubate at 70°C for 10 min to yield a homogeneous solution. Note: Briefly centrifuge the 1.5 ml microcentrifuge tube to remove drops from the inside of the lid.
- Add 200 μl ethanol (96-100%), and mix thoroughly by vortex.
- Pipet the mixture into the Spin Column CR2 (in a 2 ml collection tube) and centrifuge at 12,000 rpm (~13,400 × g) for 30 s. Discard flow-through and place the Spin Column CR2 into the collection tube.
- Add 500 μl Buffer GD to Spin Column CR2 (Ensure that ethanol is added to Buffer GD before use), and centrifuge at 12,000 rpm (~13,400 × g) for 30 s, then discard the flow-through and place the spin column into the collection tube.
- Add 600 μl Buffer PW to Spin Column CR2 (Ensure that ethanol is added to Buffer PW before use), and centrifuge at 12,000 rpm (~13,400 × g) for 30 s. Discard the flow-through and place the spin column into the collection tube.
- Repeat Step 14.
- Centrifuge at 12,000 rpm (~13,400 × g) for 2 min to dry the membrane completely.
- Place the Spin Column CR2 in a new clean 1.5 ml microcentrifuge tube, and pipet 50 μl Buffer directly to the center of the membrane. Incubate at room temperature for 2-5 min, and then centrifuge for 2 min at 12,000 rpm (~13,400 × g). Collect the elution.
