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RBC Bioscience MagCore HF48 User Manual page 37

Magcore automated nucleic acid extractor. magcore nucleic acid extraction kits
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Cultured Yeast Protocol
• Additional requirements:
• Preparation of Sorbitol Buffer:
Sample preparation
1. Harvest 3ml yeast cells (up to 5x10
remove any remaining media by aspiration.
2. Resuspend the cell pellet in 600μl sorbitol buffer (not provided).
Cell Lysis
1. Add 200U Lyticase or Zymolase (not provided). Incubate at 30°C for 30 minutes. Centrifuge the mixture for 10 min at 2,000 x g
to harvest Spheroplast.
2. Remove the supernatant and add 400μl of GT Buffer to the tube and vortex or pipette to resuspend the cell pellet.
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 5 min to
get clear tissue solution in the collection tube.
5. Pipette 400μl of clear tissue solution to the MagCore Sample Tube.
6. Place the 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®.
Sorbitol Buffer, Lyticase or Zymolase, Microcentrifuge tube.
1.2 M sorbitol, 10mM CaCl2, 0.1M Tris-Cl pH 7.5. Sterilize by filtration and store at 4°C
7
cells) by centrifugation at 5000 x g for 10 minutes. Discard the supernatant and carefully
MagCore®
34

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