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RBC Bioscience MagCore HF48 User Manual page 19

Magcore automated nucleic acid extractor. magcore nucleic acid extraction kits
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Running Time:
44 min (sample volume: 200 µl, up to 5 x 10
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. Ensure PBS buffer have been prepared for resuspend cell pellet.
Protocol
Sample Preparation
A. Cells grown in suspension
Cells grown in suspension(up to 5 x 10
min. at 300 x g in a 1.5 ml microcentrifuge tube (not provided). Remove the supernatant completely and discard, Continue with
MagCore® Operation step 1.
B. Cells grown in a monolayer
Cells grown in a monolayer(up to 5 x 10
trypsinization or using a cell scraper.
To trypsinize cells:
Determine the number of cells. Aspirate the medium and wash cells with PBS (not provided). Aspirate the PBS, and add
0.10–0.25% trypsin. After cells have detached from the dish or flask, collect them in medium, and transfer the appropriate
number of cells(up to 5 x 10
supernatant completely and discard, taking care not to disturb the cell pellet. Continue with MagCore® Operation step 1.
Using a cell scraper:
Detach cells from the dish or flask. Transfer the appropriate number of cells(up to 5 x 10
and centrifuge for 5 min. at 300 x g. Remove the supernatant completely and discard, taking care not to disturb the cell pellet.
Continue with MagCore® Operation step 1.
MagCore® Operation
1. Resuspend cell pellet with PBS Buffer to a final volume of 200 μl.
2. Transfer cell mixture 200 μl and add 20 μl Proteinase K into the MagCore Sample Tubes.
3. Place the prepared Sample Tube into well 4 of T-rack.
4. Put the Elution Tube into well 1 of T-Rack and the Tip Plus Holder Set into well 2 of T-Rack.
5. Run Code.110 program at MagCore®.
Amniotic Fluid Protocol
Sample preparation
1. Harvest cells from 10~15 ml amniotic fluid of 16~18 weeks by centrifugation for 10 minutes at 3000 rpm and
discard the supernatant.
2. Add 200µl GT Buffer (not provided) to the tube and resuspend the cell pellet, then transfer mixture to new
microcentrifuge tube.
3. Add 5~10µl ProteinaseK (10mg/ml) to the sample. Vortex for 5 seconds to mix sample.
4. Incubate at 56°C for 10 minutes until the sample lysate is clear. During incubation, invert the tube every 3
minutes.
5. Spin down the sample and apply for MagCore® HF16.
6
cells). Determine the number of cells. Centrifuge the appropriate number of cells for 5
6
cells). Cells grown in a monolayer can be detached from the culture flask by either
6
cells) to a 1.5 ml microcentrifuge tube (not provided). Centrifuge for 5 min. at 300 x g. Remove the
6
cells)
6
cells) to a 1.5 ml microcentrifuge tube
MagCore®
16

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