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make_fdr_table ERROR, No FIRE score threshold has an FDR < 0.05 #60

Description

@suyanan

I run FIRE got this error:

[Thu Jun 25 10:34:56 2026]
localrule fdr_table:
    input: temp/h3/shuffle/v0.1-shuffled-pileup.bed.gz, results/h3/additional-outputs-v0.1/coverage/h3-v0.1-minimum-coverage.txt, results/h3/additional-outputs-v0.1/coverage/h3-v0.1-maximum-coverage.txt
    output: results/h3/additional-outputs-v0.1/fire-peaks/h3-v0.1-fire-score-to-fdr.tbl
    jobid: 11
    reason: Missing output files: results/h3/additional-outputs-v0.1/fire-peaks/h3-v0.1-fire-score-to-fdr.tbl; Input files updated by another job: temp/h3/shuffle/v0.1-shuffled-pileup.bed.gz, results/h3/additional-outputs-v0.1/coverage/h3-v0.1-maximum-coverage.txt, results/h3/additional-outputs-v0.1/coverage/h3-v0.1-minimum-coverage.txt
    wildcards: sm=h3, v=v0.1
    resources: tmpdir=/DATA3/users/suyanan/research/fiberseq/analysis/20260507_1_peanut/output/run_FIRE/tmp, mem_mb=94208, mem_mib=89844

Activating conda environment: .snakemake/conda/fc1fdba7ebe99668a597ba84c09bfdc0_
[INFO][Time elapsed (ms) 501]: Header of the pileup file:
['#chrom', 'start', 'end', 'coverage', 'fire_coverage', 'score', 'coverage_shuffled', 'fire_coverage_shuffled', 'score_shuffled']
[INFO][Time elapsed (ms) 501]: Schema overrides for the pileup file:
{'column_1': String, 'column_6': Float64, 'column_9': Float64}
[INFO][Time elapsed (ms) 1318]: Done reading pileup file:
shape: (4_040_967, 9)
┌────────┬───────────┬───────────┬──────────┬───┬───────────┬────────────┬────────────┬────────────┐
│ #chrom ┆ start     ┆ end       ┆ coverage ┆ … ┆ score     ┆ coverage_s ┆ fire_cover ┆ score_shuf │
│ ---    ┆ ---       ┆ ---       ┆ ---      ┆   ┆ ---       ┆ huffled    ┆ age_shuffl ┆ fled       │
│ str    ┆ i64       ┆ i64       ┆ i64      ┆   ┆ f64       ┆ ---        ┆ ed         ┆ ---        │
│        ┆           ┆           ┆          ┆   ┆           ┆ i64        ┆ ---        ┆ f64        │
│        ┆           ┆           ┆          ┆   ┆           ┆            ┆ i64        ┆            │
╞════════╪═══════════╪═══════════╪══════════╪═══╪═══════════╪════════════╪════════════╪════════════╡
│ chr1   ┆ 83868     ┆ 83893     ┆ 2        ┆ … ┆ -1.0      ┆ 0          ┆ 0          ┆ -1.0       │
│ chr1   ┆ 83893     ┆ 84285     ┆ 2        ┆ … ┆ 33.628685 ┆ 0          ┆ 0          ┆ -1.0       │
│ chr1   ┆ 84285     ┆ 84315     ┆ 3        ┆ … ┆ 22.419123 ┆ 0          ┆ 0          ┆ -1.0       │
│ chr1   ┆ 84315     ┆ 84422     ┆ 3        ┆ … ┆ -1.0      ┆ 0          ┆ 0          ┆ -1.0       │
│ chr1   ┆ 84422     ┆ 84454     ┆ 3        ┆ … ┆ -1.0      ┆ 1          ┆ 0          ┆ -1.0       │
│ …      ┆ …         ┆ …         ┆ …        ┆ … ┆ …         ┆ …          ┆ …          ┆ …          │
│ chr1   ┆ 112416304 ┆ 112416322 ┆ 1        ┆ … ┆ -1.0      ┆ 3          ┆ 1          ┆ 21.789316  │
│ chr1   ┆ 112416322 ┆ 112416344 ┆ 1        ┆ … ┆ -1.0      ┆ 2          ┆ 1          ┆ 32.683975  │
│ chr1   ┆ 112416344 ┆ 112416654 ┆ 1        ┆ … ┆ -1.0      ┆ 2          ┆ 0          ┆ -1.0       │
│ chr1   ┆ 112416654 ┆ 112416675 ┆ 0        ┆ … ┆ -1.0      ┆ 2          ┆ 0          ┆ -1.0       │
│ chr1   ┆ 112416675 ┆ 112416884 ┆ 0        ┆ … ┆ -1.0      ┆ 1          ┆ 0          ┆ -1.0       │
└────────┴───────────┴───────────┴──────────┴───┴───────────┴────────────┴────────────┴────────────┘
/home/suyanan/.cache/snakemake/snakemake/source-cache/runtime-cache/tmpxk9ji2hw/file/home/suyanan/software/anaconda3/envs/snakemake/FIRE-main/workflow/rules/../scripts/fdr-table.py:170: DeprecationWarning: `DataFrame.melt` is deprecated; use `DataFrame.unpivot` instead, with `index` instead of `id_vars` and `on` instead of `value_vars`
  pileup.melt(
[INFO][Time elapsed (ms) 2104]: Number of Mbp in each category:
shape: (2, 2)
┌─────────┬────────────┐
│ is_real ┆ Mbp        │
│ ---     ┆ ---        │
│ bool    ┆ f64        │
╞═════════╪════════════╡
│ false   ┆ 111.002786 │
│ true    ┆ 111.002786 │
└─────────┴────────────┘
[INFO][Time elapsed (ms) 2104]: Done aggregating pileup file:
shape: (227_670, 3)
┌───────────┬─────────┬──────────┐
│ score     ┆ is_real ┆ bp       │
│ ---       ┆ ---     ┆ ---      │
│ f64       ┆ bool    ┆ i64      │
╞═══════════╪═════════╪══════════╡
│ 58.856922 ┆ true    ┆ 81       │
│ 56.8301   ┆ true    ┆ 101      │
│ 54.49021  ┆ true    ┆ 16       │
│ 54.233547 ┆ false   ┆ 101      │
│ 53.639194 ┆ false   ┆ 141      │
│ …         ┆ …       ┆ …        │
│ 1.4461327 ┆ true    ┆ 269      │
│ 1.4139963 ┆ false   ┆ 78       │
│ 1.4139963 ┆ true    ┆ 40       │
│ -1.0      ┆ false   ┆ 11864608 │
│ -1.0      ┆ true    ┆ 21596502 │
└───────────┴─────────┴──────────┘
[INFO][Time elapsed (ms) 2446]: FDR results
      threshold       FDR  shuffled_bp     real_bp
2329      -1.00  1.000000          1.0         1.0
2328       7.81  0.999879   38407185.0  38411852.0
2327       7.82  0.999997   38338210.0  38338319.0
2326       7.83  0.999168   38288191.0  38320059.0
2325       7.84  0.999017   38229169.0  38266791.0
...         ...       ...          ...         ...
4         42.47  0.162946        365.0      2240.0
3         44.53  0.244142        323.0      1323.0
2         45.93  0.321893        272.0       845.0
1         46.51  0.296569        242.0       816.0
0         54.23  0.510101        101.0       198.0

[2330 rows x 4 columns]
Traceback (most recent call last):
  File "/home/suyanan/.cache/snakemake/snakemake/source-cache/runtime-cache/tmpxk9ji2hw/file/home/suyanan/software/anaconda3/envs/snakemake/FIRE-main/workflow/rules/../scripts/fdr-table.py", line 338, in <module>
    defopt.run(main, show_types=True, version="0.0.1")
  File "/DATA3/users/suyanan/research/fiberseq/analysis/20260507_1_peanut/output/run_FIRE/.snakemake/conda/fc1fdba7ebe99668a597ba84c09bfdc0_/lib/python3.12/site-packages/defopt.py", line 308, in run
    return bind(
           ^^^^^
  File "/home/suyanan/.cache/snakemake/snakemake/source-cache/runtime-cache/tmpxk9ji2hw/file/home/suyanan/software/anaconda3/envs/snakemake/FIRE-main/workflow/rules/../scripts/fdr-table.py", line 331, in main
    fdr_table = make_fdr_table(
                ^^^^^^^^^^^^^^^
  File "/home/suyanan/.cache/snakemake/snakemake/source-cache/runtime-cache/tmpxk9ji2hw/file/home/suyanan/software/anaconda3/envs/snakemake/FIRE-main/workflow/rules/../scripts/fdr-table.py", line 197, in make_fdr_table
    raise ValueError(
ValueError: No FIRE score threshold has an FDR < 0.05. Check the input Fiber-seq data with the QC pipeline and make sure you are using WGS Fiber-seq data.
[Thu Jun 25 10:35:00 2026]
Error in rule fdr_table:
    jobid: 11
    input: temp/h3/shuffle/v0.1-shuffled-pileup.bed.gz, results/h3/additional-outputs-v0.1/coverage/h3-v0.1-minimum-coverage.txt, results/h3/additional-outputs-v0.1/coverage/h3-v0.1-maximum-coverage.txt
    output: results/h3/additional-outputs-v0.1/fire-peaks/h3-v0.1-fire-score-to-fdr.tbl
    conda-env: /DATA3/users/suyanan/research/fiberseq/analysis/20260507_1_peanut/output/run_FIRE/.snakemake/conda/fc1fdba7ebe99668a597ba84c09bfdc0_
    shell:
        
        MIN=$(cat results/h3/additional-outputs-v0.1/coverage/h3-v0.1-minimum-coverage.txt)
        MAX=$(cat results/h3/additional-outputs-v0.1/coverage/h3-v0.1-maximum-coverage.txt)
        python /home/suyanan/.cache/snakemake/snakemake/source-cache/runtime-cache/tmpxk9ji2hw/file/home/suyanan/software/anaconda3/envs/snakemake/FIRE-main/workflow/rules/../scripts/fdr-table.py -v 1 temp/h3/shuffle/v0.1-shuffled-pileup.bed.gz results/h3/additional-outputs-v0.1/fire-peaks/h3-v0.1-fire-score-to-fdr.tbl --max-cov $MAX --min-cov $MIN
        
        (one of the commands exited with non-zero exit code; note that snakemake uses bash strict mode!)

Removing output files of failed job fdr_table since they might be corrupted:
results/h3/additional-outputs-v0.1/fire-peaks/h3-v0.1-fire-score-to-fdr.tbl
[Thu Jun 25 10:38:38 2026]
Finished job 33.
19 of 39 steps (49%) done
Removing temporary output temp/h3/decorate/v0.1-chr1.dec.bed.gz.
Shutting down, this might take some time.
Exiting because a job execution failed. Look above for error message
Complete log: .snakemake/log/2026-06-25T102041.210767.snakemake.log
WorkflowError:
At least one job did not complete successfully.

make_fdr_table step, this Error No FIRE score threshold has an FDR < 0.05. Check the input Fiber-seq data with the QC pipeline and make sure you are using WGS Fiber-seq data.

How can I debug this from QC pipeline, my peanut sample is m6A mark rate 4.29% with depth 32.23.

Look forward to your reply, thanks.

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