DNA-only protocol
The expected concentration after the first PCR ranges from 0.2 – 2.0 ng/μL.
Low concentrations may be caused by:
- A low cell number input during encapsulation (< 3,000 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
- Forgot the UV-light treatment of emulsions after Barcoding – Tapestri Instruments with serial numbers MBT-2019 or lower only
- Used the wrong UV light during UV-light treatment of emulsions after Barcoding – Tapestri Instruments with serial numbers MBT-2019 or lower only
High concentrations may be caused by:
- A high cell number input during encapsulation (> 4,000 cells/μL)
- Excessive drop merging after Barcoding (amplification takes place in ‘quasi-bulk’ environment)
- Excessive Barcoding Bead loading after Barcoding