Automated gDNA extraction using the Felix

This protocol uses a magnetic bead-based whole nucleic acid extraction kit run on the Felix. It can take either 200 µL or 400 µL of sample as input, eluting in 30 µL or 50 µL.

It’s not recommended to elute in 30 µL when running the 400 µL sample input protocol, as the volumes of elution solution and beads are too close and may result in an inefficient elution.

96-, 48-, 24-, and 12-well plates can easily be harvested in 200 µL or less, and 6-well plates can easily be harvested in 400 µL.

 

200 µL sample input

Dispense reagents only to the wells you will be using. Partial plates can be reused for future DNA extractions. If your sample is not 200 µL, make it up to 200 µL by adding PBS (or similar).

  1. Shake magnetic beads well before using. For each extraction, mix 265 µL Binding Buffer + 10 µL Magnetic Beads. Make at least a 10% excess. Dispense 275 µL Binding Buffer + Magnetic Beads per well of a 96-well deep well plate.

  2. Dispense 500 µL Wash Buffer per well of a 96-well deep well plate.

  3. Dispense 800 µL 80% ethanol per well of a 96-well deep well plate.

  4. Dispense 30 or 50 µL Elution Solution per well of a 96-well hard-shell PCR plate.

  5. Dispense 5 uL proteinase K per well of a 96-well deep well plate, then add 200 µL of sample.

  6. Open the protocol NYUExtract-MVPII_200uL_v2.2_(elution_30/elution_50).bms from the MagMAX folder on the Felix computer, and click the green ‘Play’ button. You will be prompted with the deck layouts, load your paltes accordingly.

    • Use CyBi-Tips 1000 µL (filtered), which you will have to manually move from their tip trays to the black tip racks on OL3317-11-105 stands. This can be made easier by using a 200 µL Gilson multichannel pipette to move the tips.

  7. When the protocol is complete, remove the elution plate, now containing your eluted DNA. If the plates were not filled completely, they can be saved for future DNA extractions. Make sure you indicate which wells have been used (although it should be pretty clear based on the waste products they now contain). If the plates have now been completely used, discard them.

 

400 µL sample input

Dispense reagents only to the wells you will be using. Partial plates can be reused for future DNA extractions. If your sample is not 400 µL, make it up to 400 µL by adding PBS (or similar).

  1. Shake magnetic beads well before using. For each extraction, mix 530 µL Binding Buffer + 20 µL Magnetic Beads. Make at least a 10% excess. Dispense 550 µL Binding Buffer + Magnetic Beads per well of a 96-well deep well plate.

  2. Dispense 800 µL Wash Buffer per well of a 96-well deep well plate.

  3. Dispense 1,500 µL 80% ethanol per well of a 96-well deep well plate.

  4. Dispense 30 or 50 µL Elution Solution per well of a 96-well hard-shell PCR plate.

  5. Dispense 10 µL proteinase K per well of a 96-well deep well plate, then add 400 µL of sample.

  6. Open the protocol NYUExtract-MVPII_400uL_v2.2_(elution_30/elution_50).bms from the MagMAX folder on the Felix computer, and click the green ‘Play’ button. You will be prompted with the deck layouts, load your paltes accordingly.

    • Use CyBi-Tips 1000 µL (filtered), which you will have to manually move from their tip trays to the black tip racks on OL3317-11-105 stands. This can be made easier by using a 200 µL Gilson multichannel pipette to move the tips.

  7. When the protocol is complete, remove the elution plate, now containing your eluted DNA. If the plates were not filled completely, they can be saved for future DNA extractions. Make sure you indicate which wells have been used (although it should be pretty clear based on the waste products they now contain). If the plates have now been completely used, discard them.