Library / Automated NGS Library Prep / Steps

Protocol steps

The full workflow, hands-free. Volumes and timings are illustrative — match them to your kit.

1

Normalize input

The robot reads your input concentrations and dilutes each well to a uniform target (e.g. 20 ng in 30 µL), so every sample enters fragmentation on equal footing.

~10 minVision confirms tip pickup and well-by-well dispense.
2

Fragment / tagment

Fragmentation or tagmentation mix is added and the plate is moved to the thermocycler for the timed enzymatic step. Fragment size is set by incubation time/temperature per your kit.

15–30 min
3

End repair & A-tailing

End-repair and A-tailing enzymes are added directly and incubated, preparing fragments for adapter ligation.

~30 min
4

Adapter ligation

Adapters and ligation mix are added; the robot mixes gently and incubates to ligate adapters onto both fragment ends.

~15 min
5

Post-ligation SPRI cleanup

SPRI beads remove enzymes and adapter dimers. The magnet engages, supernatant is removed, two 80% ethanol washes follow, beads air-dry, and DNA is eluted.

~20 minVision verifies magnet engagement, supernatant clearance, and that beads aren't over-dried.
6

Indexing PCR

Unique dual indexes and PCR master mix are added per well; the plate cycles on the thermocycler for the minimum cycles needed to preserve complexity.

~30 min
7

Dual-sided size selection

A two-ratio SPRI selection removes both large fragments and small adapter dimers, leaving the target insert range. The bead ratios are set to your desired size window.

~25 minVision checks each bead transfer and wash.
8

Final elution

Final libraries are eluted into a clean plate, ready for quantification, the Bioanalyzer/TapeStation, and pooling.

~5 min
Hands-on touchpoints: loading reagents, moving the plate to/from the thermocycler if it's off-deck, and sealing for PCR. Everything between is walk-away.