Home FAQs When using the Monarch Spin High-Capacity RNA Cleanup Kit (3 mg) (NEB #T2060), is it necessary to use a thermal mixer for elution?

FAQ: When using the Monarch Spin High-Capacity RNA Cleanup Kit (3 mg) (NEB #T2060), is it necessary to use a thermal mixer for elution?

For most samples with RNA size ≤ 5 kb, incubation at room temperature without thermal mixing will not impact RNA recovery if two or more elutions are performed. If a thermal mixer is unavailable, the recommendations below can be followed to maximize recovery:

Temperature recommendation: Nuclease-free water can be pre-warmed to 37°C before adding to the column for elution. Alternatively, after adding nuclease-free water, the column (placed in a user-provided RNase-free microfuge tube) can be incubated in a dry air incubator at 37°C.

Mixing recommendation: Do not vortex. Gentle rocking on a rocking platform can be used during incubation. Alternatively, after the incubation period, gentle mixing by carefully pipetting the nuclease-free water in the column several times over the membrane layer can enhance recovery. Take care not to touch the membrane layer with the pipette tip.