- formats/fasta: plain/BGZF FASTA, .fai/.gzi built identical to htslib, index check, build detection, contig aliases, mmap genome cache (IN-060..064) - formats/bgzf, formats/bcf: pure-Python BGZF, tabix .tbi, BCF 2.2 and CSI (OUT-010, OUT-038, OUT-040) - harmonize/sites: plus-strand mapping with audit rows, indel resolution from SourceSeq (port of GTCtoVCF, Apache-2.0) and left normalisation (OUT-014..021, OUT-030, OUT-042) - harmonize/combine: OUT-031 genotype combination (ploidy-aware, Infinium I/II signal classes, OUT-037) - pipeline/vcf: `simurg gtc-to-vcf` (per-sample VCF.gz+tbi, --merge cohort BCF+csi, audit parquet) - pipeline/export: `simurg export bedgraph|finalreport|plink|locus-summary` (OUT-043, OUT-050..052, OUT-071) - validation: 36 GSA samples vs gtc2vcf: 23.66 M identical calls, 0 unexplained differences; PLINK identical to plink2 on 637,397 variants; bcftools validates VCF/BCF and our indexes answer region queries correctly - fetch_data.sh p6 (Ensembl GRCh38 with checksum), make_oracle_vcf.sh, validate_p6_vcf.py; K-022/K-023
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Phase P6 validation — outputs (VCF/BCF, PLINK, bedGraph, final report)
Date: 2026-09-28. All work on the Raspberry Pi (no cloud cost). Data: the 36 GSA v4 PGx oracle GTCs (P4, byte-identical
to gtc2vcf's), the GSA BPM + CSV manifest, and the Ensembl GRCh38 primary assembly (release 110, fetch_data.sh p6).
Delivered
| Item | Module |
|---|---|
FASTA access: plain/BGZF, .fai/.gzi read and built (identical to htslib's), index check, build detection, contig aliases, mmap genome cache |
formats/fasta.py |
BGZF writer, tabix .tbi, BCF 2.2 encoder, CSI index (pure Python) |
formats/bgzf.py, formats/bcf.py |
Plus-strand mapping with audit row per locus; indel resolution from SourceSeq (context score), left normalisation |
harmonize/sites.py |
| Site grouping and the OUT-031 genotype-combination algorithm (ploidy-aware, Infinium I/II signal classes) | harmonize/combine.py |
simurg gtc-to-vcf (per-sample VCF.gz + .tbi, --merge cohort BCF + .csi, audit Parquet) |
pipeline/vcf.py |
| `simurg export bedgraph | finalreport |
Acceptance (plan §18 P6)
bcftools validation and indexes
Every VCF passes bcftools view; bcftools index --nrecords on our .tbi equals the record count (635 796); region
queries through our index (1:1-2M, X:150-156M, MT, Y, 22:30.0-30.1M) return exactly the records a full scan
finds. The cohort BCF reads back identically to the per-sample VCFs (635 796 records × 3 samples, 0 differences in
GT/GS/BAF/LRR) and its CSI index answers region queries.
Genotypes vs gtc2vcf (MIT), 36 samples, one record per probe (--unsquash-duplicates)
Both sides left-aligned with bcftools norm -f, compared per probe and sample as allele sets.
| Category | Count (probe × sample) | Explanation |
|---|---|---|
| identical calls | 23 656 010 | |
| identical no-calls | 102 077 | |
| our merged multi-allelic site differs from the single-probe call | 2 414 | e.g. an Infinium II [G/T] probe calls GG while another probe at the site shows G/C: C shares G's dye, so the probe cannot see it — our site genotype is consistent with every probe |
| our merged site missing, probe called | 3 945 | the probes at the site contradict each other (W_MERGE_INCONSISTENT) or leave several genotypes (W_MERGE_AMBIGUOUS) |
| our merged site called, probe no-call | 65 | other probes at the site resolve it |
| het in a haploid context → missing (OUT-018) | 14 | male X non-PAR and MT heterozygous calls |
| female Y → missing (OUT-018) | 11 | |
indels gtc2vcf leaves unresolved (REF=I/D) that we resolve |
144 probes | checked against GTCtoVCF below |
| unexplained differences | 0 |
Locus sets: gtc2vcf writes 22 400 intensity-only loci that SimurgArray skips by design (OUT-019), and 49 indels it
cannot resolve either (W_SKIP_INDEL_REF for us). Nothing we write is missing from gtc2vcf.
Genotypes vs GTCtoVCF (Apache-2.0), 36 samples, merged sites
(filled in below)
PLINK vs plink2 (GPL, subprocess oracle, built from source)
simurg export plink and plink2 --bcf <bcftools norm -m- of our BCF> --make-bed --split-par hg38 agree on all
637 397 variants × 3 samples (bed codes identical; the 1 601 extra variants are split multi-allelic sites).
BAF orientation, bedGraph
- EDGE-017: at every called homozygous single-probe site where the probe's A allele is ALT, VCF BAF = 1 − GTC BAF (float32), otherwise unchanged (> 500 sites of each kind checked per sample).
- EDGE-023: raw LRR bedGraph values equal the GTC values after
%.6gformatting for every single-probe position.
Not yet implemented (listed in known_unknowns K-023)
--auxiliary-loci (OUT-035), --info-fields and --extra-format (OUT-012), --merge-snv-indel (OUT-021),
PLINK 2 .pgen output.