FAQ: Why do I need to incubate my column for 5 minutes with elution buffer (nuclease-free water)?

Elution of large amounts of RNA (> 50 µg) can be maximized by incubating the elution buffer (nuclease-free water) with the column for 5 minutes at room temperature prior to spinning. Decreasing the incubation time will decrease RNA recovery.


RNA recovery is increased by incubating the column with the elution buffer (nuclease-free water) prior to the elution spin

rRNA (16S and 23S Ribosomal Standard from E.coli, Sigma) was purified using the Monarch RNA Cleanup Kit (500 µg, NEB #T2050). 100 µl of nuclease-free water was added to the column and incubated for either 0,1,3 or 5 minutes at room temperature before spinning to elute the RNA. The percent recovery of RNA was calculated from the resulting A260 as measured using a Trinean Dropsense 16. A five-minute incubation period produced the maximum yield.