Clean-up using AMPure XP beads
Mix the reagent well so that the reagent appears homogeneous and consistent in color.
Add 117 μL of homogenous AMPure XP beads to each adaptor ligated DNA sample (in either 1.5 mL LoBind tubes or 0.2 mL LoBind tubes).
Mix well by pipetting up and down at least 10 times.
Incubate at room temperature for 5 min.
Put the tube in the magnetic stand and wait for the solution to clear (which should take approximately 3–5 min).
Keep the tube in the magnetic stand.
Do not touch the beads while you carefully discard the cleared solution from the tubes.
Continue to keep the tube in the magnetic stand/rack whilst adding 500 μL (1.5 mL LoBind tubes) or 200 μL (0.2mL LobBind tubes) of 70% ethanol to each tube.
(wash 1/2).
Let the tube sit for 1 min to allow any disturbed beads to settle, and remove the ethanol.
(wash 1/2).
Continue to keep the tube in the magnetic stand/rack whilst adding 500 μL (1.5 mL LoBind tubes) or 200 μL (0.2mL LobBind tubes) of 70% ethanol to each tube.
(wash 2/2).
Let the tube sit for 1 min to allow any disturbed beads to settle, and remove the ethanol.
(wash 2/2).
Seal the tube or plate and centrifuge briefly (260 x g for 30 sec).
Return the tube to the magnetic stand/rack and wait 1 min.
Remove any remaining ethanol using a P20 pipette and tip, being careful not to touch the bead pellet.
Dry the samples on a 37 °C heat block for 3–5 min or until the residual ethanol completely evaporates.
IMPORTANT: Do not over-dry as this will decrease yield.
Add 32 μL nuclease-free water directly to the bead pellet, mix well by pipetting up and down at least 10 times.
Incubate for 3 min at room temperature.
Centrifuge briefly to consolidate the sample and place on a magnetic stand/rack for 2–3 min or until the solution is clear.
Remove 30 μL of the supernatant and transfer to a fresh LoBind tube.
The beads can be discarded at this time.
