Bio-Rad Aurum™ Total RNA Fatty and Fibrous Tissue Kit - Manual (Page 28)

14. Add 700 µl of high stringency wash solution to the RNA binding
column. Centrifuge for 30 sec at >12,000 x g. Discard the high
stringency wash solution from the wash tube and place the column back
into the same wash tube.
15. Add 700 µl of low stringency wash solution (already
supplemented with ethanol) to the RNA binding column.
Centrifuge for 1 min at >12,000 x g. Discard the filtrate from the wash tube
and place the column back into the same wash tube.
16. Centrifuge for an additional 2 min at >12,000 x g to remove
residual wash solution.
17. Transfer the RNA binding column to a 1.5 ml capped
microcentrifuge tube (provided).
18. Pipet 40 µl (or 30 µl) † of the elution solution onto the center of
the membrane at the bottom of the RNA binding column.
† Note: When isolating total RNA from small amounts of starting material
(<10 mg of tissue or 500,000 cells), perform a single elution with 30 µl of
the elution solution. Do not perform step 21.
19.Incubate 1 min for complete soaking and saturation of the
membrane.
20. Centrifuge for 2 min at >12,000 x g to elute the total RNA.
21. Repeat steps 18 and 19 using 40 µl of the elution solution if the
starting amounts of starting material is more than 10 mg of
tissue or 500,000 cells.
Note: The eluted total RNA samples can be used immediately in downstream
applications. Alternatively, the RNA sample can be aliquoted and stored at
–20°C or –70°C for later use.
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