What this protocol does
This protocol automates a seqWell-style high-plex library prep on the Undergrad: tagmentation-based fragmentation, indexing PCR, plate pooling, and SPRI cleanup — designed for many low-input samples at once. It targets the labs that batch hundreds of libraries, where manual prep is both the bottleneck and the biggest source of batch effects.
Who it's for
Microbial genomics, plasmid QC, and high-plex sequencing applications that prep libraries in bulk and need reproducibility across large plates.
From intent to atomic steps
Layer 1 · Intent — plain languageDescribe the automated seqwell library prep the way you would brief a labmate — no scripting, no deck programming.
2 Protocol stages3 Robot actions4 Atomic actions — vision-verified
1Normalize input~15 min
Dilute each well to the kit's target input so all samples tagment equivalently (see the Plate Normalization protocol).
◉Vision: Vision confirms per-well dispense.
2Tagment~20 min
Add tagmentation mix and incubate to simultaneously fragment and tag the DNA.
3Index PCR~40 min
Add unique dual indexes and master mix; amplify on the thermocycler for the minimum cycles needed.
4Pool~15 min
Combine indexed libraries into a single pool per plate.
◉Vision: Vision verifies each pooling transfer.
5SPRI cleanup~25 min
Bead-clean the pool: magnet, two 80% ethanol washes, brief dry, and elute.
◉Vision: Vision verifies magnet engagement, washes, and that beads aren't over-dried.
6Elute~5 min
Elute the final pooled library, ready for QC and sequencing.