Take one 0.2 ml PCR tube and two 1.5 ml Eppendorf DNA LoBind tubes. Label them as follows:
- 0.2 ml PCR tube: label as "Tube 1"
- 1.5 ml Eppendorf DNA LoBind tube: label as "Tube 2"
- 1.5 ml Eppendorf DNA LoBind tube: label as "Tube 3"

Transfer 2 µl of Lambda DNA (LMD) to Tube 1.

Add 8 µl nuclease-free water to Tube 1.

Add 1 µl Fragmentation Mix (FRA) to Tube 1.

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 30°C for two minutes, then at 80°C for two minutes.
• Take the tube out from the thermal cycler or heat block, spin down briefly in a microfuge, and put the tube on ice for approximately 30-60 seconds to cool it down.

Pipette 1.5 μl of Rapid Adapter (RA) into Tube 2.

Add 3.5 μl of Adapter Buffer (ADB) to Tube 2.

Mix the contents of Tube 2 by pipetting up and down. Spin down briefly in a microfuge.

Add 1 μl of the Tube 2 contents into Tube 1.

• Mix the contents of the tube by flicking the tube with your finger
• Spin down briefly in a microfuge
• Incubate for 5 minutes at room temperature

Store the library on ice until you are ready to load it into the flow cell.