Instruments
- Undergrad with the Accuris AutoMATE 96 for liquid handling
- Magnetic module for SPRI bead cleanups and size selection
- Thermocycler for fragmentation/tagmentation and indexing PCR (on-deck or adjacent)
- Onboard cameras enabled for vision verification of bead steps (default)
Input requirements
Before the run, confirm input meets your kit's spec. The robot will normalize concentration, but it can't fix degraded or contaminated input.
| Parameter | Typical target |
|---|---|
| Input amount | 10–100 ng DNA (kit-dependent; low-input kits go lower) |
| Purity | A260/280 ≈ 1.8; A260/230 ≈ 2.0 |
| Quantification | Fluorometric (e.g. Qubit), not just absorbance |
| Integrity | High-molecular-weight for fragmentation kits; assess by gel/TapeStation |
Labware
- Input/sample plate (normalized) and a destination PCR plate
- Index plate (unique dual indexes recommended)
- Reagent reservoirs / cold block for enzymes
- SPRI bead reservoir, fresh 80% ethanol trough, elution-buffer reservoir
- Filter tips (multiple racks — fresh tips per transfer)
Reagents (kit-agnostic)
- Fragmentation/tagmentation mix, end-repair & A-tailing enzymes
- Adapters and ligation mix
- Unique dual indexes + indexing PCR master mix
- SPRI magnetic beads, freshly prepared 80% ethanol, elution buffer (EB/Tris)
Deck layout
Keep enzymes on the cold block, place SPRI beads and ethanol adjacent to the magnetic module to minimize travel, and load index and reagent reservoirs in workflow order. The Undergrad confirms each position by vision before starting, so placement is forgiving — but workflow-order loading gives the cleanest run.
Make fresh 80% ethanol the morning of the run. Evaporated/old ethanol is the single most common cause of poor SPRI cleanups and low library yield — see Troubleshooting.