FAQ: What is the smallest volume of nuclease-free water that can be used for elution with the Monarch RNA Cleanup Columns?

T2030/T2037: 6 µl; typical elution volumes are 6-20 µl

T2040/T2047: 20 µl; typical elution volumes are 20-50 µl

T2050/T2057: 50 µl; typical elution volumes are 50-100 µl

Using lower elution volumes will likely result in variance due to inconsistent wetting of the matrix. Yield may slightly increase if a larger volume is used, but the RNA will be less concentrated.

*A second elution can be performed to maximize RNA recovery. This second elution can be performed using a fresh aliquot of nuclease-free water or the eluent from the first elution to maximize recovery while minimizing volume.

Recovery of RNA from Monarch RNA Cleanup Kits with Varying Elution Volumes



rRNA (10, 50 or 500 µg, respectively of 16S and 23S Ribosomal Standard from E. coli, Sigma) was purified using a Monarch RNA Cleanup Kit (10 µg, NEB #T2030) (50 µg, NEB #T2040) (500 µg, NEB #T2050). Nuclease-free water was used to elute the RNA. The percent recovery of the RNA was calculated from the resulting A260 as measured using a Trinean DropSense 16. ~80% of RNA can be efficiently recovered in 6 µl from the Monarch RNA Cleanup Kit (10 µg, NEB #T2030), 20 µl from the Monarch RNA Cleanup Kit (50 µg, NEB #T2040), and 50 µl from the Monarch RNA Cleanup Kit (500 µg, NEB #T2050).