OpenPlant Automation Protocols
DNA extraction of 96-well cultures with magnetic beads
Description
This protocol carries out DNA extraction of bacterial cultures in a 96-deepwell plate with the magnetic module
For help with how to run Jupyter Notebook protocols, refer to our tutorial
Labware and Liquids
- Right mount: An 8-channel p300 pipette
- Slot 1: 12-well reservoir with buffers (fill up to volume for required samples: max 96 samples)
- Well 1: Lysis buffer (Up to 12 mL)
- Well 2, 3: Neutralization buffer (Up to 15 mL each well)
- Well 4, 5: Wash buffer (Up to 15 mL each well)
- Well 6, 7: Ethanol (Up to 12 mL each well)
- Well 8: Elution Buffer (Up to 10mL)
- Slot 7: 96-deepwell plate (square wells) with ressuspended bacterial cell pellets (in 120 uL ressuspension buffer)
- Slot 9: Output plate in 96-well format
- Slot 10: Magnetic module with 96-deepwell plate (square wells) with 100uL of magnetic beads
- Slot 11: 1-well reservoir for buffer waste
- Slots 1, 2, 3, 5, 6: Tipracks for p300
Execution
- Start by centrifuging the culture plate, removing the supernatant and ressuspending the pellets in ressuspension buffer
- Set up the deck as described above
- Run the cells one by one, paying attention to prompts and instructions
Script
DNA extraction of 96-well cultures with magnetic beads
Tips and best practices
This is a protocol with many manual steps. You will have to monitor and work along the robot throughout the whole protocol
The addition of ethanol after the neutralization step helps to precipitate nucleotides and enhances the final yield, but increases the risk of RNA contamination. Using fresh RNAse on the Ressuspension Buffer helps, but still expect some RNA content.