Low concentrations may be caused by:
- A low cell number input during encapsulation (< 2,800 cells/μL)
- A clog during cell barcoding that prevented Barcoding Beads from entering the cartridge
- Used expired reagents
- PCR settings were incorrect – Use the correct ramp rate, temperatures, and annealing/extension times
- Used the incorrect Ampure XP reagent/sample ratio
- The sample and Ampure XP reagent were not at room temperature before clean up
- 80% EtOH was not freshly prepared
High concentrations may be caused by:
- A high cell number input during encapsulation (> 3,200 cells/μL)
- Excessive drop merging after Barcoding (amplification takes place in ‘quasi-bulk’ environment)