RBC Bioscience MagCore HF48 User Manual page 29

Running Time:
33min(sample volume: 400 µl)
Preparation before using
The kit procedures are optimized for a maximum of 100 mg of wet-weight or 20 mg of dried starting material.
Exceeding the recommended maximum amount of starting material will result in inefficient lysis, resulting in low yield and
purity.
Tissue Dissociation Protocol
1. Cut 50 mg (up to 100 mg) of fresh or frozen plant tissue or 5 mg (up to 20 mg) of dried sample.
2. Grind the sample with mortar and pestle under liquid nitrogen to a fine powder. For some plant samples, liquid nitrogen may
be unnecessary for homogenization.
3. Transfer it into a microcentrifuge tube (not provided).
Lysis Step:
1. Add 400μl GP1 Buffer and 5μl RNase A (10mg/ml) into the microcentrifuge tube and mix by vortexing. Do not mix GP1 Buffer
with RNase A before use.
2. Incubate at 65°C for 10 minutes. During incubation, invert the tube every 5 minutes.
3. Add 100μl GP2 Buffer and mix by vortexing.
4. Incubate on ice for 3 minutes. Place a Filter Column into a 2 ml Collection Tube and apply the entire lysate from previous step to
the Filter Column.
5. Centrifuge for 3 minutes at full speed (13,000 rpm).
6. Discard the Filter Column and carefully transfer clarified lysate(about 400μl) in the collection Tube to the MagCore Sample
Tubes.
7. Place the prepared Sample Tube into well 4 of T-rack.
8. Put the Elution Tube into well 1 of T-Rack and the Tip Plus Holder Set into well 2 of T-Rack.
9. Run Code.301 program at MagCore®.
Fungal Tissue Protocol
Sample preparation
1. Collect the fungal tissue up to 20 mg.
2. Grind the sample with mortar and pestle under liquid nitrogen to a fine powder.
3. Transfer it into a microcentrifuge tube (not provided). Do not allow the sample to thaw.
Cell Lysis
1. Add 400μl GP1 Buffer and 5μl RNase A (10mg/ml) into the microcentrifuge tube and mix by vortexing. Do not mix GP1 Buffer
with RNase A before use.
2. Incubate at 65°C for 10 minutes. During incubation, invert the tube every 5 minutes.
3. Add 100μl GP2 Buffer and mix by vortexing.
4. Incubate on ice for 3 minutes. Place a Filter Column into a 2 ml Collection Tube and apply the entire lysate from previous step to
the Filter Column.
5. Centrifuge for 3 minutes at full speed (13,000 rpm).
6. Discard the Filter Column and carefully transfer clarified lysate(about 400μl) in the collection Tube to the MagCore Sample
Tubes.
7. Place the prepared Sample Tube into well 4 of T-rack.
8. Put the Elution Tube into well 1 of T-Rack and the Tip Plus Holder Set into well 2 of T-Rack.
9. Run Code.301 program at MagCore®.
MagCore®
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