The two main sources impacting emulsions quality/stability are:
- Thermal cycling
- Emulsion handling
- Incorrect PCR tubes used other than the below validated tubes
- 0.2 mL Axygen MAXYmum Recovery PCR tubes (Axygen, PCR-02-L-C or PCR-02D-L-C)
- 0.2 mL TempAssure PCR Flex-Free 8-Tube Strips (North America: USA Scient., 1402-4700, color: natural)
- 0.2 mL 8-Strip Non-Flex PCR Tubes (Europe: Starlab, I1402-3600, color: natural)
- Total volume of emulsion + oil greater than or less than ~100 uL
- 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 email@example.com.
- 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 User Guides for instructions on the safe-handling of emulsions.