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40
RNeasy Mini Handbook 10/2019
10. Add 500 µl Buffer RPE to each RNeasy spin column.
Note: Buffer RPE is supplied as a concentrate. Ensure that ethanol is added to Buffer RPE
before use (see “Things to do before starting”).
11. Switch on the vacuum. Apply vacuum until transfer is complete. Switch off the vacuum
and ventilate the vacuum manifold.
The flow-through is collected in the QIAvac 24 Plus, the QIAvac 24 base, or the QIAvac
6S waste tray.
12. Add 500 µl Buffer RPE to each RNeasy spin column.
13. Switch on the vacuum. Apply vacuum until transfer is complete. Switch off the vacuum
and ventilate the vacuum manifold.
The flow-through is collected in the QIAvac 24 Plus, the QIAvac 24 base, or the QIAvac
6S waste tray.
14. Remove the RNeasy spin columns from the vacuum manifold, and place each in a 2 ml
collection tube (supplied). Close the lids gently, and centrifuge at full speed for 1 min.
15. Place each RNeasy spin column in a new 1.5 ml collection tube (supplied). Add 30–
50 µl RNAse-free water directly to each spin column membrane. Close the lids gently,
and centrifuge for 1 min at ≥8000 x
g
(≥10,000 rpm) to elute the RNA.
16. If the expected RNA yield is >30 µg, repeat step 15 using another 30–50 µl RNAse-free
water, or using the eluate from step 15 (if high RNA concentration is required). Reuse the
collection tubes from step 15.
If using the eluate from step 15, the RNA yield will be 15–30% less than that obtained
using a second volume of RNAse-free water, but the final RNA concentration will be
higher.
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