Low fluorescence
| Possible Cause |
Solution |
| Degraded MasterMix |
Store at -20 °C, thaw at 2 - 8 °C no longer than 30 min. Vortex and short spin before use. |
| Unverified cycler |
Use verified cycler. List of verified cyclers here. |
| Incorrect consumables |
Use recommended consumables. List of consumables in device manual. |
| Clear PCR strips on BioRad CFX96 |
Use white PCR strips on BioRad CFX96 |
| Incorrect transport medium |
Use TE buffer or Nuclease-free water. |
| Air bubbles in PCR mix solution |
Spin PCR strip before PCR. |
| Photosensitive MasterMix |
Minimize exposure of the MasterMix to light during handling to avoid loss of fluorescent signal intensity. |
|
Combination of parameters with unequal fluorescence on Rotor-Gene Q
|
Place the positive control from the problematic PCR kit in well 1 and enable Auto-Gain Optimization, or run the problematic kit in a separate experiment.
|
No fluorescence
| Possible cause |
Solution |
| Incorrect PCR setup |
Contact support. Templates can be provided. |
| Incorrect consumables |
Use recommended consumables. List of consumables in device manual. |
| Incorrect transport medium |
Use TE buffer or Nuclease-free water. |
| Photosensitive MasterMix |
Minimize exposure of the MasterMix to light during handling to avoid loss of fluorescent signal intensity. |
| Degraded PCR reagents |
Transport the PCR kit on dry ice. |
|
Combination of parameters with unequal fluorescence on Rotor-Gene Q
|
Place the positive control from the problematic PCR kit in well 1 and enable Auto-Gain Optimization, or run the problematic kit in a separate experiment.
|
All
