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RBC Bioscience MagCore HF48 User Manual page 17

Magcore automated nucleic acid extractor. magcore nucleic acid extraction kits
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Running Time:
44 min(sample volume :200 µl) // 57 min(sample volume :400 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.
Whole Blood Protocol
1. Take a new Sample Tube and add 20μl of Proteinase K (10mg/ml) to 200μl of equilibrated whole blood sample.
(40μl Proteinase K to 400μl whole blood).
2. Place the prepared Sample Tube into well 4 of T-Rack.
3. Put the Elution Tube into well 1 of T-Rack and the Tip Plus Holder Set into well 2 of T-Rack.
4. Run Code.106 program at MagCore®.
Optional Step: RNA Degradation
If RNA-Free genomic DNA is required, perform these optional steps before adding Proteinase K.
1. Add 4μl RNase A(50mg/ml; not provided) into the sample lysate.
2. Incubate the sample at room temperature for 20min
Buffy Coat modify Protocol
RBC Lysis Buffer:
150mM NH
Cl , 10mM KHCO
4
Buffy Coat Preparation by RBC Lysis
1. Take 600 ~ 700μl whole blood into 2ml microcentrifuge tube.
Don't take more than 700μl whole blood sample; it will cause the leakage situation during process.
2. Add 1ml RBC Lysis Buffer and mix the buffer and whole blood sample by upside down.
3. Shake the mixture, 100rpm 5mins.
4. Centrifuge the mixture 13,000rpm 1 min.
5. Discard supernatant.
6. Repeat step 2 ~ step 5 to wash the sample again.
7. Add 400μl RBC Lysis Buffer and 20μl proteinase K to resuspend the pellet and transfer into MagCore Sample Tube.
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.106 program at MagCore®.
Buffy Coat Preparation by Centrifugation
1. Take 2 ~ 5ml whole blood sample and centrifuge at 1,500rpm 10mins.
2. Use plastic drop to take white buffy coat layer in the middle of whole blood sample.
3. Move the buffy coat into new microcentrifuge tube.
4. Take 80 ~ 100μl buffy coat sample into MagCore Sample Tube and add RBC Lysis Buffer or PBS until 400μl then add 20μl of
proteinase K.
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.106 program at MagCore®.
Note : We suggest to select 150 ~200μl elution buffer, it can get better elution efficiency in both of these methods.
Normally the concentration is higher than 150ng/μl under such elution volume.
, 0.1mM EDTA.
3
MagCore®
14

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