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M2551 Mag-Bind® FFPE RNA 96 Kit Protocol
6.
Add 25 µL Proteinase K Solution (20 mg/mL). Incubate at 55°C for 15-30 minutes with
occasional mixing. If necessary, extend the incubation to 1-3 hours or until the tissue
is completely lysed.
7.
Incubate at 80°C for 15 minutes.
8.
Immediately centrifuge at 4,000 x g for 5 minutes. The paraffin will form a thin layer
on top of the lysate solution.
9.
Use a 1 mL pipette tip or large orifice tip to penetrate the paraffin layer, transfer 200
μL cleared lysate into a new round-well plate.
10. Add 200 µL MFB Buffer, 20 µL Mag-Bind® Particles SC, and 430 µL of 100% ethanol.
Mix thoroughly by vortexing for 20 seconds or pipetting up and down 10-20 times.
Note: If the RNA content from sample is expected low or miRNA is the target, then
add 10 µL LPA Buffer.
11. Let sit at room temperature for 5-10 minutes.
12. Place the plate onto a magnetic separation device for deep-well plates and wait 7-10
minutes or until the Mag-Bind® Particles SC are cleared from solution.
Note: If using the MSD-01 magnetic separation device, a 500 µL processing plate
(EZ960-01/02) is required for the rest of the protocol. Since the total volume of the
sample is around 850 µL, this particular magnetic separation device requires the
sample be transferred twice to process whole sample.
13. Aspirate and discard the cleared supernatant.
14. Remove the plate from the magnetic separation device.
15. Add 400 µL RNA Wash Buffer II. Resuspend the Mag-Bind® Particles SC thoroughly by
vortexing for 20 seconds or pipetting up and down 10-20 times.
Note: RNA Wash Buffer II must be diluted with 100% ethanol prior to use. Please see
instructions on Page 4.
7
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