package obiprimer import ( "cmp" "math" "slices" "sync" "git.metabarcoding.org/obitools/obitools4/obitools4/pkg/obidefault" "git.metabarcoding.org/obitools/obitools4/obitools4/pkg/obithermo" ) // ampRecord is an amplicon of a retained pair, in one sequence. type ampRecord struct { hash uint64 // the barcode: see ampHash pair int32 taxon int32 // taxon of the sequence at the chosen rank, -1 when it has none role Role } // refHit is an amplicon of a retained pair on the reference sequence. type refHit struct { up, down seqMatch swapped bool } // ampHash hashes the amplicon between the two primers read from the side of // the first primer of the pair (reverse complemented when the upstream match is // the second primer), so that identical amplicons give the same value // whatever their strand. Non ACGT bases hash as a fifth symbol. func ampHash(raw []byte, from, to int, reverse bool) uint64 { const prime = 1099511628211 h := uint64(14695981039346656037) ^ uint64(to-from) h *= prime if !reverse { for _, b := range raw[from:to] { h = (h ^ uint64(baseCode[b])) * prime } return h } for i := to - 1; i >= from; i-- { c := baseCode[raw[i]] if c < 4 { c = 3 - c } h = (h ^ uint64(c)) * prime } return h } // amplicons holds what the second phase gathers on the retained pairs. type amplicons struct { parts [][]ampRecord // one slice per worker, each sorted by pair minTm1 []float32 // by pair: lowest Tm of the first primer over the example sites minTm2 []float32 refs map[int32]refHit } // pair returns the amplicons of one retained pair, as one slice per worker. func (a *amplicons) pair(i int32) [][]ampRecord { out := make([][]ampRecord, 0, len(a.parts)) byPair := func(r ampRecord, p int32) int { return cmp.Compare(r.pair, p) } for _, part := range a.parts { lo, _ := slices.BinarySearchFunc(part, i, byPair) hi, _ := slices.BinarySearchFunc(part, i+1, byPair) if hi > lo { out = append(out, part[lo:hi]) } } return out } // collectOptions selects what collectAmplicons gathers. type collectOptions struct { examplesOnly bool // skip the counterexamples and the other sequences model *obithermo.Model // melting temperatures of the example sites (nil: none) reference bool // record the amplicons on the reference sequence } // collectAmplicons gathers, in parallel, one record per amplicon of a retained // pair, in the sequences of the corpus (all of them, or the examples only), and // the lowest melting temperatures over the example sites. The records are not // copied nor regrouped: each worker keeps its own, sorted by pair. func (ps *primerSet) collectAmplicons(c *Corpus, taxa *Taxa, p PairParams, index map[pairKey]int32, opt collectOptions) *amplicons { model := opt.model lp := p.PrimerLength workers := obidefault.ParallelWorkers() npairs := len(index) res := &licons{ parts: make([][]ampRecord, workers), minTm1: make([]float32, npairs), minTm2: make([]float32, npairs), refs: map[int32]refHit{}, // written by the single worker that reads the reference } inf := float32(math.Inf(1)) for i := range npairs { res.minTm1[i], res.minTm2[i] = inf, inf } var tmMu sync.Mutex // oriented[primer][0] is the reverse complement of the word, [1] the word itself. oriented := make([][2][]byte, len(ps.primers)) for i, pr := range ps.primers { oriented[i][0] = []byte(orientedWord(pr.Word, lp, false)) oriented[i][1] = []byte(orientedWord(pr.Word, lp, true)) } jobs := make(chan int) var wg sync.WaitGroup for w := range workers { wg.Add(1) go func() { defer wg.Done() site1, site2 := make([]byte, lp), make([]byte, lp) lowTm1, lowTm2 := map[int32]float32{}, map[int32]float32{} for s := range jobs { raw := c.Seqs[s].Sequence() n := len(raw) role := c.Roles[s] if opt.examplesOnly && role != Example { continue } ps.forEachOccurrence(c, s, p, func(o occurrence) { pi, ok := index[o.key] if !ok { return } if opt.reference && p.HasReference && s == p.Reference { if old, seen := res.refs[pi]; !seen || o.up.pos < old.up.pos { res.refs[pi] = refHit{o.up, o.down, o.swapped} } } res.parts[w] = append(res.parts[w], ampRecord{ hash: ampHash(raw, int(o.up.pos)+lp, int(o.down.pos), o.swapped), pair: pi, taxon: taxa.Of[s], role: role, }) if role != Example || model == nil { return } fp := oriented[o.up.primer][b2i(!o.up.reverse)] rp := oriented[o.down.primer][b2i(o.down.reverse)] tmF, tmR := inf, inf if windowAsIs(site1, raw, int(o.up.pos), n, p.Circular) { if t, err := model.Tm(fp, site1); err == nil && !math.IsNaN(t) { tmF = float32(t) } } if windowRevComp(site2, raw, int(o.down.pos), n, p.Circular) { if t, err := model.Tm(rp, site2); err == nil && !math.IsNaN(t) { tmR = float32(t) } } if o.swapped { tmF, tmR = tmR, tmF // slot 1 is the downstream primer } lowTm1[pi] = min(lowOr(lowTm1, pi, inf), tmF) lowTm2[pi] = min(lowOr(lowTm2, pi, inf), tmR) }) } slices.SortFunc(res.parts[w], func(a, b ampRecord) int { return cmp.Compare(a.pair, b.pair) }) tmMu.Lock() for pi, t := range lowTm1 { res.minTm1[pi] = min(res.minTm1[pi], t) } for pi, t := range lowTm2 { res.minTm2[pi] = min(res.minTm2[pi], t) } tmMu.Unlock() }() } for s := range c.Seqs { jobs <- s } close(jobs) wg.Wait() return res } func lowOr(m map[int32]float32, k int32, def float32) float32 { if v, ok := m[k]; ok { return v } return def } func b2i(b bool) int { if b { return 1 } return 0 } const upper = "ACGT" // windowAsIs copies the len(dst) bases at pos as upper-case ACGT; it reports // false if one of them is not A, C, G or T. func windowAsIs(dst, raw []byte, pos, n int, circular bool) bool { for t := range dst { i := pos + t if circular { i %= n } c := baseCode[raw[i]] if c > 3 { return false } dst[t] = upper[c] } return true } // windowRevComp writes the reverse complement of the len(dst) bases at pos. func windowRevComp(dst, raw []byte, pos, n int, circular bool) bool { l := len(dst) for t := range dst { i := pos + l - 1 - t if circular { i %= n } c := baseCode[raw[i]] if c > 3 { return false } dst[t] = upper[3-c] } return true }