RBC Bioscience MagCore HF48 User Manual page 43

Running Time:
44min (sample volume :200 µl)
Preparation before using
1. Add 1.1ml PK Storage Buffer to the Proteinase K tube and mix by vortexing. Store prepared Proteinase K (10mg/ml) at -20°C.
2. Freshly prepared 20mg/ml Lysozyme solution before use. (for Gream + bacteria isolation, Lysozyme solution is necessary)
Lysozyme ( not provided ) + Lysozyme Reaction Buffer = Lysozyme Solution
For Sputum Specimens
Specimens Decontamination
1. Fresh prepare 0.5% NALC in 2% NaOH, 1.5% Na-Citrate solution.
(Ex: 0.25g NALC in 50mL NALC-NaOH solution)
2. Mix 10mL specimen with 10mL NALC-NaOH sol'n, RT°C for 15 min.
3. Add 25mL PBS, mix and centrifuge 3000 x g for 15 min.
4. Discard supernatant, resuspend pellet with 200µl Lysozyme solution and transfer to the MagCore Sample Tube.
5. Incubate for at least 30min at 37°C. During incubation, vortex the tube every 5min.
Cell Lysis
1. Add 4μl RNase A (50mg/ml) to sample mixture(including any precipitate) and vortex to mix sample.
2. Incubate at room temperature for 10min.
3. Resuspend sample mixture by pipetting.
4. Adding 40μl Proteinase K(10mg/ml) to sample mixture and vortex to mix sample.
5. Place the prepared Sample Tube into well 4 of T-rack.
6. Put the Elution Tube into well 1 of T-Rack and the Tip Plus Holder Set into well 2 of T-Rack.
7. Run Code.502 program at MagCore®.
General Protocol
1. Harvest bacteria (maximum 5x 10
Discard supernatant.
2. Resuspend bacterial pellet in 200μl Lysozyme Solution by vortexing or pipetting. (if target is Gram- bacteria, please use
Lysozyme Reaction Buffer )
3. Incubate for at least 30min at 37°C and vortex the tube every 5min. (for Gram- bacteria isolation, you can skip this step)
4. Add 4μl RNase A (50mg/ml) to sample mixture(including any precipitate) and vortex to mix sample.
5. Incubate at room temperature for 10min.
6. Resuspend sample mixture by pipetting.
7. Adding 40μl Proteinase K(10mg/ml) to sample mixture and vortex to mix sample.
8. Place the prepared Sample Tube into well 4 of T-rack.
9. Put the Elution Tube into well 1 of T-Rack and the Tip Plus Holder Set into well 2 of T-Rack.
10. Run Code.502 program at MagCore®.
6
cells) into the MagCore Sample Tube by centrifuging at 5000 x g(8000rpm) for 3min.
MagCore®
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