Low cfDNA Recovery from Incomplete Elution

Symptom

Final DNA concentration is lower than expected despite successful binding and washing. A significant portion of DNA remains bound to the beads after the elution step.

1. 1 Insufficient mixing of beads with Nuclease-free Water during elution
2. 2 Not all beads resuspended during the elution step, with visible clumps remaining
3. 3 Elution incubation time shorter than the required 5 minutes
4. 4 Absence of thermal mixer/shaker during the 5-minute elution incubation

1. 1 Pipette mix Nuclease-free Water and beads thoroughly until all beads are completely resuspended
2. 2 Visually confirm no bead clumps remain before proceeding to incubation
3. 3 Incubate resuspended beads for full 5 minutes on a thermal mixer/shaker
4. 4 If thermal mixer unavailable, perform additional pipette mixing after the 5-minute incubation

Related Video (1)

Thermo Fisher Scientific ★ 85

How to Isolate Cell-Free DNA from Plasma | cfDNA Purification

"Directly demonstrates the complete cfDNA isolation workflow including the critical elution step where incomplete mixing causes DNA loss from beads"