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Mag-Bind® FFPE RNA 96 Protocol with Xylene
6.
Centrifuge at 4,000 x g at room temperature for 5 minutes to pellet the tissue.
Note: If the tissue does not form a tight pellet, centrifuge for an additional 3 minutes.
7.
Carefully remove and discard the xylene without disturbing the pellets.
8.
Add 1 mL 100% ethanol to each well. Mix thoroughly by vortexing for 20 seconds.
9.
Centrifuge at 4,000 x g for 5 minutes to pellet the tissue samples. The pellets should
appear opaque.
10. Carefully remove and discard the ethanol. Remove any liquid drops with a pipette.
11. Repeat Steps 8-10 for a second ethanol wash step.
12. Air dry the tissue pellet for 10-20 minutes.
Note: It is critical to completely dry the sample before the Proteinase K digestion
step. Ethanol residue will effect the efficiency of the Proteinase K digestion. If a
vacuum oven is available, place the tube into the vacuum oven preset at 45°C for
10-30 minutes.
13. Add 250 μL RML Buffer and 25 μL Proteinase K Solution (20 mg/mL). Resuspend the
pellet by vortexing or pipetting up and down 20 times.
14. Incubate at 55°C for 15 minutes.
15. Incubate at 80°C for 15 minutes.
16. Centrifuge at 4,000 x g at room temperature for 5 minutes.
17. Carefully transfer 200 μL cleared supernatant into a new round-well plate.
11
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