Min NucleoSpin RNA Clean-up, User Manual

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15 MACHEREY-NAGEL – 05 / 2014, Rev. 05
NucleoSpin ® RNA Clean-up
5 Wash and dry silica membrane
1 st wash
Add 250 μL Buffer RA2 to the NucleoSpin ® RNA Clean-
up Column. Centrifuge for 30 s at 8,000 x g. Discard
flow-through and reuse Collection Tube
+ 250 μL RA2
8,000 x g ,
30 s
2
nd
wash
Add 700 μL Buffer RA3 to the NucleoSpin
®
RNA Clean-
up Column. Centrifuge for 30 s at 8,000 x g. Discard
flow-through and reuse Collection Tube.
+ 700 μL RA3
8,000 x g ,
30 s
3
rd
wash
Add 350 μL Buffer RA3 to the NucleoSpin
®
RNA Binding
Column. Centrifuge for 2 min at 8,000 x g .
Transfer the NucleoSpin
®
RNA Clean-up Column to a
nuclease-free Collection Tube (1.5 mL, supplied). Open
the lid of the column and let the membrane dry for 3 min.
If for any reason, the liquid level in the Collection Tube
has reached the NucleoSpin ® RNA Clean-up Column after
centrifugation, discard flow-through and centrifuge again.
The procedure ensures complete removal of ethanol from
the column.
+ 350 μL RA3
8,000 x g ,
2 min
6 Elute RNA
Elute the RNA in 60 μL RNase-free H
2
O, (supplied) and
immediately centrifuge at 8,000 x g . for 1 min.
If higher RNA concentrations are desired, elution can be
done with 40 μL. Overall yield, however, will decrease when
using smaller volumes.
For further alternative elution procedures see section 2.4.
+ 60 μL
RNase-free
H
2
O
8,000 x g ,
1 min