RBC Bioscience MagCore HF48 User Manual page 32

For Solid Animal Tissue
Additional Requirements: Microcentrifuge Tube.
1. Cut the solid tissue to small pieces (up to 30 mg) and put into a microcentrifuge tube.
2. Add 400μl GT Buffer and 20μl Proteinase K(10mg/ml) to the tube and mix by vortexing.
3. Incubate at 55°C for 90min until the sample has been completely lysed.
4. If there are any insoluble residues in the tube, transfer the supernatant to Filter Column and centrifuge at full speed for 5min to
get clear tissue solution in the Collection Tube.
5. Pipette 400μl of clear tissue solution to the MagCore Sample Tube.
6. Place the prepared Sample Tube into well 4 of T-rack.
7. Put the Elution Tube into well 1 of T-Rack and the Tip Plus Holder Set into well 2 of T-Rack.
8. Run Code.401 program at MagCore®.
For Stool Sample
Additional Requirements: Microcentrifuge Tube.
1. Weight 180-200mg stool in a 2ml microcentrifuge tube and place on ice. If the sample is liquid, pipet 200μl into microcentrifuge
tube. Cut the end of pipet tip to make pipetting easier. If the sample is frozen, use a scalpel or spatula to scrape bits of stool into
microcentrifuge tube on ice.
2. Add 1.5ml GT Buffer to sample. Vortex continuously for 1 min or until the stool sample is thoroughly homogenized. This is very
important to vortex sample thoroughly to ensure maximum DNA concentration in the final elutes.
3. Incubate the suspension for 5 min at 70°C. This step can increase DNA recovery 3-5 fold, if the sample target is Gram-positive
bacteria, please increase to 95°C for cells lysis.
4. Vortex for 15 seconds and centrifuge sample at full speed (13,000rpm) for 1 min to pellet stool particles.
5. Pipet 400μl of the supernatant into a new 1.5ml microcentrifuge tube.
6. Add 20μl Proteinase K (10mg/ml) to the sample mixture and vortex to mix. Incubate at 60°C for 2~3 hours to lyse the sample.
7. If there are any insoluble residues in the tube, transfer the supernatant to Filter Column and centrifuge at full speed for 5min to
get clear tissue solution in the Collection Tube.
8. Pipette 400μl of clear tissue solution to the MagCore Sample Tube.
9. Place the prepared Sample Tube into well 4 of T-rack.
10. Put the Elution Tube into well 1 of T-Rack and the Tip Plus Holder Set into well 2 of T-Rack.
11. Run Code.401 program at MagCore®.
www.rbcbioscience.com
29