Bead clean-up (single tube)

Prepare the following reagents/materials:

or Homemade SPRI beads

Fresh 70% Ethanol

Nuclease-free water or elution buffer

DNA sample/PCR product

Clean 1.5 mL tubes

Magnetic tube rack

In a 1.5 mL tube, add resuspended beads to sample/PCR product in ratio specified in main protocol.

Flick tube gently to mix beads and sample.

Allow mixture to incubate for 0h 5m 0s at Room temperature

Place tube on a magnetic rack.

Equipment

Value	Label
Magnetic Stand	NAME
Magnetic Stand	TYPE
Thermo Scientific	BRAND
MR02	SKU
Any magnetic rack that fits your tubes will suffice.	SPECIFICATIONS

Wait ~1-3 minutes for the beads and buffer to separate. Beads will stick to magnetic side of the tube and the solution should be clear.

Using a pipettor, remove and discard the clear supernatant solution. DNA will remain in the tube bound to the beads.

Add 500 μL 70% EtOH to tube

• Do not disturb the beads! Pipette into the opposite side of the tube.
  Note

  Volume of 70% EtOH can be adjusted. Just so long as the amount added is enough to cover beads.

Incubate at Room temperature for 0h 0m 30s

Using a pipettor, remove and discard the EtOH. Be careful not to disturb or aspirate the beads.

Repeat the Step 4 EtOH wash. Try to remove as much EtOH as possible without disturbing the beads.

Leaving tube cap open, allow beads to dry for a maximum of 90 seconds.

Remove tube from magnetic rack and resuspend DNA/beads by adding a minimum of 15 μL nuclease-free water or desired buffer.

Flick tube gently to resuspend beads in elution.

Incubate at Room temperature for 0h 2m 0s- 0h 5m 0s

Place tube back on magnetic rack. Allow beads and elution to separate for 0h 1m 0s .

Slowly aspirate out clear supernatant, now containing DNA, and transfer to final clean container.