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Instruction Manual
7.
Once the user confirms that the MCV Volume is correct, Stabilization Duration begins
8.
A prompt to confirm that Media Exchange phase is complete will display.
Press
to acknowledge that Media Exchange Phase is complete.
8.2.7 Transfection Phase (Process-dependent)
This procedure shows how to add transfection complex to the MCV via one of the two AseptiQuik® G connectors on the MCV headplate.
In this phase, the recirculation pump and control loops are automatically stopped so that the transfection reagent is mixed in the MCV for
a specified amount of time.
CAUTION:
Care should be taken with all components that contact the bioreactor after adding the transfection reagent. Adhere to the
biosafety protocols.
Follow the manufacturer’s protocol for transfection:
• Concentration of DNA in media (µg/mL)
• Depending on the transfection method, the amount of transfection reagent in media may need to be adjusted
• Use endotoxin-free DNA
• Adjust the volume of transfection reagent added to the MCV depending on protocol.
1. Prepare transfection reagent.
Fill a bottle with transfection complex in the biosafety cabinet. The bottle should have an AseptiQuik® G connection.
2. Connect bottle.
Pick one of the two AseptiQuik® G connectors from the MCV headplate to connect the bottle
containing the transfection complex.
3.
Enter setpoints:
- Transfection Volume (mL)
- Agitation Speed (rpm)
- Mixing Duration (min.)
- Stabilization Delay Duration (min.)
- Recirculation Ramp Duration (min.)
Press
to start.
NOTE:
The Recirculation Ramp Duration is the ramp time after stabilization from minimum
recirculation pump flow rate to the flow rate preceding Transfection.
4.
A prompt will remind the user to complete setting up the transfection bottle and connecting
lines.
Press
when complete.