Thaw the following reagents at room temperature, and then keep on ice:

• Lambda DNA (LMD)
• Ligation Adapter (LA)
• DNA Control Strand (DCS)
• Ligation Buffer (LNB)
• Long Fragment Buffer (LFB)
• AMPure XP Beads (AXP)
• Elution Buffer (EB)
• Sequencing Buffer (SB)
• Library Beads (LIB)
• Flow Cell Tether (FCT)
• Flow Cell Flush (FCF)

Once the contents of each tube have thawed, mix the Lambda DNA (LMD), DNA Control Strand (DCS) and Ligation Buffer (LNB) by pipetting up and down and vortex the other reagents. Spin down all the thawed reagents briefly in a microfuge.

Program a thermal cycler to run at 20°C for 5 minutes, then 65°C for 5 minutes. Alternatively, set two heat blocks at 20°C and 65°C.