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7 changes: 7 additions & 0 deletions new-test-data/README.md
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# Pixelator 0.29 test data overlay

Updated fixtures for pixelator 0.29.0 live under `new-test-data/pna/`.
Unchanged files are copied from `testdata/pna/`; changed or new files are stored here.

The v2 samplesheet with the prerelease panel is at
`new-test-data/samplesheet/pna/samplesheet_proxiome_v2.csv`.
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## Test datasets origin

This test data is generated by Pixelgen Technologies AB for the nf-core/pixelator pipeline.

Synthetic PNA test data in this folder was generated with internal prototype scripts.

### FASTQ generation procedure

Data generation used deterministic seeds for reproducibility.

Generation was performed in two steps:

1. Build a synthetic edgelist parquet with:

- 20 cells
- 600 nodes per cell
- 22200 edges in total (200 crossing edges)
- 20% of antibodies are hashing antibodies

2. Convert each parquets into paired FASTQ files and using the corresponding PNA panel definitions:

- proxiome-v1-immuno-155-v1.1 panel for `sample1`, `sample1_reseq`, `sample2`
- proxiome-v2-immuno-155-prerelease panel for `pool1`, `pool1_reseq`, `pool2`

This generates the files in `testdata/pna/fastq`, including:

- `sample1*_R1.fastq.gz`, `sample1*_R2.fastq.gz`, `sample2_R1.fastq.gz`, `sample2_R2.fastq.gz`
- `pool1*_R1.fastq.gz`, `pool1*_R2.fastq.gz`, `pool2_R1.fastq.gz`, `pool2_R2.fastq.gz`
- corresponding `*.metadata.json` files

### Module output generation (`testdata/pna/modules`)

Module artifacts were then generated with Pixelator `single-cell-pna` subcommands, in this order:

1. `amplicon`
2. `demux`
3. `collapse` + `combine-collapse`
4. `graph`
5. `denoise` (pool-level for proxiome-v2)
6. `sample-calling`
7. `layout`
8. `analysis`

Representative options (from module metadata):

- `demux`: `--design proxiome-v2 --panel proxiome-v2-immuno-155-prerelease`
- `sample-calling`: `--samplesheet samplesheet_proxiome_v2.csv --enrichment-threshold 10.0`
- `graph`: `--component-size-min-threshold 100 --multiplet-recovery`
- `layout`: `--layout-algorithm coarsened_pmds_3d`
- `analysis`: `--compute-proximity --proximity-nbr-of-permutations 100`
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