DNA tagmentation

Thaw kit components at Room temperature , spin down briefly using a microfuge and mix by pipetting as indicated below:

• Lambda DNA (50 μg/ml): thaw at RT, briefly spin down, mix well by pipetting
• Fragmentation Mix (FRA): not frozen, briefly spin down, mix well by pipetting
• Rapid Adapter (RAP): not frozen, briefly spin down, mix well by pipetting
• Sequencing Buffer (SQB): thaw at RT, briefly spin down, mix well by pipetting*
• Loading Beads (LB): thaw at RT, briefly spin down, mix by pipetting or vortexing immediately before use
• Sequencing Tether (SQT): thaw at RT, briefly spin down, mix well by pipetting

Prepare the DNA in Nuclease-free water

Transfer ~200ng genomic DNA into a 1.5 ml Eppendorf DNA LoBind tube

Adjust the volume to 3.75µL with Nuclease-free water

Mix by flicking the tube to avoid unwanted shearing

Spin down briefly in a microfuge

In a 0.2 ml thin-walled PCR tube, mix the following:

3.75µL 200 ng template DNA

1.25µL FRA

Mix gently by flicking the tube, and spin down.

Incubate the tube at 30°C for 0h 1m 0s and then at80°C for 0h 1m 0s . Briefly put the tube on ice to cool it down.