The two main sources impacting emulsions quality/stability are:
- Thermal cycling
- Emulsion handling
Thermal cycling
- Incorrect PCR tubes used, other than the 0.2 mL Axygen MAXYmum Recovery PCR Tubes and PCR-02-L-C
- PCR tubes are not fully closed
- No skirt or supporting ‘corner’ PCR tubes used for even heat transfer
A layer of fluid above the emulsion layer after the targeted PCR (1st PCR) is indicative of a failure mode.
If you notice an aqueous layer on top of the emulsion layers after thermal cycling, contact support@missionbio.com.
Emulsion handling
- Static electricity on tubes, gloves, or devices
- Mechanical stress on tubes
Emulsion tubes are very sensitive to uneven heating, physical stress, or harsh handling. Please refer to the Tapestri Single-Cell DNA Sequencing V2 User Guide for instructions on the safe-handling of emulsions.