Transfer 20 µl of Lambda DNA (LMD) to a clean 0.2 ml PCR tube (Tube 1).

Add 27 µl nuclease-free water to the tube.

Add 1 µl DNA CS (DCS) to the tube.

Add 3.5 µl of the NEBNext FFPE DNA Repair Buffer to the tube.

Add 2 µl of the NEBNext FFPE DNA Repair Mix to the tube.

Add 3.5 µl of the Ultra II End-prep reaction buffer to the tube.

Add 3 µl of the Ultra II End-prep enzyme mix to the tube.

Mix the contents of the tube by flicking the tube with your finger. Spin down briefly in a microfuge.

• Incubate in heat blocks or a thermal cycler at 20°C for 5 minutes, then at 65°C for 5 minutes
• Take the tube out from the thermal cycler or heat block, and spin down briefly in a microfuge

Transfer the Lambda DNA sample (Tube 1) to a clean 1.5 ml Eppendorf DNA LoBind tube (Tube 2).

Add 60 µl of resuspended AXP to Tube 2 with the Lambda DNA sample, and mix by pipetting up and down.

Put the tube in a Hula mixer, and leave to incubate for 5 minutes.

While the DNA sample is incubating, transfer 350 µl of pure ethanol to a clean 1.5 ml DNA LoBind tube.

Add 150 µl nuclease-free water to the ethanol.

Mix the contents of the tube by flicking the tube with your finger. Spin down briefly in a microfuge.

Take Tube 2 with the Lambda DNA sample off the Hula mixer, and spin down in a microfuge.

Place Tube 2 in a magnetic rack, and wait for the beads to collect in a pellet near the magnet, and the solution to become clear.

Keep the tube on the magnet to pipette off and discard the supernatant. Take care to not disturb the pellet.

Add 200 µl of the freshly-prepared ethanol to Tube 2, without taking the tube off the magnetic rack or disturbing the pellet.

Ensure the pellet is covered with ethanol, then pipette off and discard the ethanol.

Repeat the previous step:
Add 200 µl of the freshly-prepared ethanol to the tube, without taking the tube off the magnetic rack or disturbing the pellet.

Ensure the pellet is covered with ethanol, then pipette off and discard the ethanol.

Take the tube off the magnetic rack and spin down briefly in a microfuge.

Place Tube 2 back in the magnetic rack, and wait for the beads to collect in a pellet near the magnet. There may be a small amount of residual liquid at the bottom of the tube. Pipette off and discard this liquid.

Remove the tube from the magnetic rack, add 60 µl nuclease-free water to the tube, and resuspend the beads by flicking the tube. Leave the tube to incubate for 2 minutes at room temperature.

Place the tube back in the magnetic rack, and wait for the beads to collect in a pellet near the magnet.

Pipette off the 60 µl of eluate and transfer it to a clean 1.5 ml Eppendorf DNA LoBind tube (Tube 3). Discard Tube 2 with the pellet.