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 .
