Monarch Spin Columns S2C were designed without a retaining ring in order to prevent contamination from buffer carryover and ensure maximum purity of eluted DNA. When the protocol is carried out as specified, the membranes are held in place and should not dislodge. However, the membrane can expand and dislodge under very specific conditions:
- The column is loaded more than once
- The lysed sample or the wash buffer is allowed to sit on the membrane for too long
- The column is vortexed
- The input amounts are exceeded significantly
- The membrane could be more likely to dislodge during low-speed spins. For kits containing orange-tinted Monarch Spin Columns S2C (NEB #T3017), follow the current protocols, which do not use a low-speed spin (1,000 x g for 3 minutes)
Accordingly, we recommend that the protocol is followed as specified and that the column is never reloaded or vortexed.
