diff --git a/charcoal/Snakefile b/charcoal/Snakefile index 2693ed8..cc3a62d 100644 --- a/charcoal/Snakefile +++ b/charcoal/Snakefile @@ -204,7 +204,7 @@ class Checkpoint_HitListPairs: # wait for the results of 'hitlist_make_contigs_matches'; # this will trigger exception until that rule has been run. - checkpoints.hitlist_make_contigs_matches.get(g=g) + checkpoints.hitlist_make_contigs_matches_all.get(g=g) match_accs = get_hitlist_match_accs(g) p = expand(self.pattern, g=g, acc=match_accs) @@ -230,12 +230,12 @@ rule all: @toplevel rule clean: input: - expand(output_dir + '/{f}.clean.fa.gz', f=genome_list) + expand(output_dir + '/{g}.clean.fa.gz', g=genome_list) # CTB: alias for 'clean'; remove later after making sure it's not in docs :) rule all_clean_contigs: input: - expand(output_dir + '/{f}.clean.fa.gz', f=genome_list) + expand(output_dir + '/{g}.clean.fa.gz', g=genome_list) # check config files only @toplevel @@ -294,11 +294,11 @@ def make_moltype_compute_args(moltype): # generate a signature for a query genome # CTB TODO: update for sourmash sketch -rule contigs_sig: +rule contigs_sig_wc: input: - genome_dir + '/{filename}' + genome_dir + '/{g}' output: - stage1_dir + '/{filename}.sig' + stage1_dir + '/{g}.sig' conda: 'conf/env-sourmash.yml' params: scaled = config['scaled'], @@ -309,15 +309,16 @@ rule contigs_sig: {params.moltype} {input} -o {output} """ + + # run a search, query.x.database. -rule prefetch_all: +rule prefetch_all_matches_wc: input: - query = stage1_dir + '/{filename}.sig', + query = stage1_dir + '/{g}.sig', databases = config['gather_db'] output: - csv = stage1_dir + '/{filename}.matches.csv', - matches = stage1_dir + '/{filename}.matches.zip', - txt = stage1_dir + '/{filename}.matches.txt' + csv = stage1_dir + '/{g}.matches.csv', + txt = stage1_dir + '/{g}.matches.txt' params: moltype = "--{}".format(config['moltype'].lower()), gather_scaled = config['gather_scaled'], @@ -326,22 +327,26 @@ rule prefetch_all: shell: """ sourmash prefetch {input.query} {input.databases} -o {output.csv} \ {params.moltype} --scaled {params.gather_scaled} \ - --save-matches {output.matches} \ --threshold-bp {params.threshold_bp} >& {output.txt} cat {output.txt} - touch {output.csv} {output.matches} + touch {output.csv} """ -# generate contigs taxonomy -rule make_contigs_taxonomy_json: +@toplevel +rule prefetch_all_matches: input: - genome = genome_dir + '/{f}', - genome_sig = stage1_dir + '/{f}.sig', - matches = stage1_dir + '/{f}.matches.zip', + expand(stage1_dir + '/{g}.matches.csv', g=genome_list) - lineages = config['lineages_csv'] +# generate contigs taxonomy +rule make_contigs_search_taxonomy_wc: + input: + genome = genome_dir + '/{g}', + genome_sig = stage1_dir + '/{g}.sig', + matches_csv = stage1_dir + '/{g}.matches.csv', + lineages = config['lineages_csv'], + databases = config['gather_db'] output: - json = stage1_dir + '/{f}.contigs-tax.json', + json = stage1_dir + '/{g}.contigs-tax.json', conda: 'conf/env-sourmash.yml' params: match_rank = default_match_rank, @@ -349,20 +354,27 @@ rule make_contigs_taxonomy_json: python -m charcoal.contigs_search_taxonomy \ --genome {input.genome} --lineages-csv {input.lineages} \ --genome-sig {input.genome_sig} \ - --matches-sig {input.matches} \ + --matches-csv {input.matches_csv} \ + --databases {input.databases} \ --json-out {output.json} \ --match-rank {params.match_rank} """ +@toplevel +rule make_contigs_search_taxonomy: + input: + expand(stage1_dir + '/{g}.contigs-tax.json', g=genome_list) + # compare taxonomy for contigs in a genome; this generates hit list and # genome summary outputs. -rule compare_taxonomy_single: +rule compare_taxonomy_single_wc: input: json = stage1_dir + '/{g}.contigs-tax.json', - sig = stage1_dir + '/{g}.matches.zip', + matches_csv = stage1_dir + '/{g}.matches.csv', lineages = config['lineages_csv'], provided_lineages = provided_lineages_file, - genome_list = genome_list_file + databases = config['gather_db'], + genome_list = genome_list_file, output: hit_list_csv = stage1_dir + '/{g}.hitlist_for_filtering.csv', summary_csv = stage1_dir + '/{g}.genome_summary.csv', @@ -384,7 +396,8 @@ rule compare_taxonomy_single: --min_f_ident={params.min_f_ident} \ --min_f_major={params.min_f_major} \ --match-rank={params.match_rank} \ - {wildcards.g} + {wildcards.g} \ + --databases {input.databases} """ # combine all of the individual hit lists into a single hitlist summary file. @@ -466,11 +479,12 @@ rule make_hitlist_matches_info_csv: # generates list of contaminant & non-contaminant accessions for genomes # on the hitlist. -checkpoint hitlist_make_contigs_matches: +checkpoint hitlist_make_contigs_matches_all: input: genome = genome_dir + '/{g}', genome_sig = stage1_dir + '/{g}.sig', - matches = stage1_dir + '/{g}.matches.zip', + matches_csv = stage1_dir + '/{g}.matches.csv', + databases = config['gather_db'], lineages = config['lineages_csv'], hitlist = output_dir + '/stage1_hitlist.csv' output: @@ -482,20 +496,26 @@ checkpoint hitlist_make_contigs_matches: python -m charcoal.contigs_list_contaminants \ --genome {input.genome} --lineages-csv {input.lineages} \ --genome-sig {input.genome_sig} \ - --matches-sig {input.matches} \ + --matches-csv {input.matches_csv} \ + --databases {input.databases} \ --hitlist {input.hitlist} \ --json-out {output.matches_json} \ --match-rank {params.match_rank} """ +@toplevel +rule hitlist_make_contigs_matches: + input: + expand(stage2_dir + '/{g}.matches.json', g=genome_list) + # run a mashmap comparison of two genomes. rule mashmap_compare: input: - query = genome_dir + '/{f}', + query = genome_dir + '/{g}', target = ancient('genbank_genomes/{acc}_genomic.fna.gz'), output: - cmpfile = stage2_dir + '/{f}.x.{acc}.mashmap.align', - outfile = stage2_dir + '/{f}.x.{acc}.mashmap.out', + cmpfile = stage2_dir + '/{g}.x.{acc}.mashmap.align', + outfile = stage2_dir + '/{g}.x.{acc}.mashmap.out', conda: 'conf/env-mashmap.yml' shell: """ mashmap -q {input.query} -r {input.target} -o {output.cmpfile} \ @@ -549,12 +569,12 @@ rule combine_postprocessing_csv: # actually do cleaning. @CTB out of date, fixme later! rule clean_contigs: input: - genome = genome_dir + '/{f}', - json = stage1_dir + '/{f}.contigs-tax.json', + genome = genome_dir + '/{g}', + json = stage1_dir + '/{g}.contigs-tax.json', hit_list = output_dir + '/stage1_hitlist.csv', output: - clean = output_dir + '/{f}.clean.fa.gz', - dirty = output_dir + '/{f}.dirty.fa.gz', + clean = output_dir + '/{g}.clean.fa.gz', + dirty = output_dir + '/{g}.dirty.fa.gz', conda: 'conf/env-sourmash.yml' shell: """ python -m charcoal.clean_genome \ diff --git a/charcoal/compare_taxonomy.py b/charcoal/compare_taxonomy.py index 56503d9..0ebc642 100644 --- a/charcoal/compare_taxonomy.py +++ b/charcoal/compare_taxonomy.py @@ -120,12 +120,23 @@ def choose_genome_lineage(guessed_genome_lineage, provided_lineage, match_rank, return genome_lineage, comment, needs_lineage -def get_genome_taxonomy(matches_filename, genome_sig_filename, provided_lineage, +def get_genome_taxonomy(matches_filename, database_list, + genome_sig_filename, provided_lineage, tax_assign, match_rank, min_f_ident, min_f_major): - try: - siglist = list(sourmash.load_file_as_signatures(matches_filename)) - except (ValueError, AssertionError) as e: - siglist = None + # load the matches from prefetch as a picklist + picklist = sourmash.picklist.SignaturePicklist('prefetch') + picklist.load(matches_filename, picklist.column_name) + + # load all of the matches in the database, as found by prefetch; + # select on them; and then aggregate into MultiIndex. + # CTB note: currently, this loads all the signatures into memory. + # Alternatively we could do something with LazyLoadedIndex maybe? + + siglist = [] + for filename in database_list: + db = sourmash.load_file_as_index(filename) + db = db.select(picklist=picklist) + siglist += list(db.signatures()) if not siglist: comment = 'no matches for this genome.' @@ -248,12 +259,13 @@ def main(args): detected_contam = {} summary_d = {} - matches_filename = os.path.join(dirname, genome_name + '.matches.zip') + matches_filename = os.path.join(dirname, genome_name + '.matches.csv') genome_sig = os.path.join(dirname, genome_name + '.sig') lineage = provided_lineages.get(genome_name, '') contigs_json = os.path.join(dirname, genome_name + '.contigs-tax.json') x = get_genome_taxonomy(matches_filename, + args.databases, genome_sig, lineage, tax_assign, match_rank, @@ -430,6 +442,8 @@ def cmdline(sys_args): p.add_argument('--min_f_ident', type=float, default=F_IDENT_THRESHOLD) p.add_argument('--min_f_major', type=float, default=F_MAJOR_THRESHOLD) p.add_argument('--match-rank', required=True) + p.add_argument('--databases', help='sourmash databases', required=True, + nargs='+') p.add_argument('genome') args = p.parse_args() diff --git a/charcoal/conf/env-reporting.yml b/charcoal/conf/env-reporting.yml index 570d9ad..1047cc2 100644 --- a/charcoal/conf/env-reporting.yml +++ b/charcoal/conf/env-reporting.yml @@ -9,7 +9,7 @@ dependencies: - notebook>=6,<7 - plotly>=4.9.0,<5 - ipykernel - - sourmash>=4.2.0,<5 + - sourmash>=4.2.2,<5 - pip - pip: - git+https://github.com/dib-lab/charcoal.git diff --git a/charcoal/conf/env-sourmash.yml b/charcoal/conf/env-sourmash.yml index 3115bfc..8d347d6 100644 --- a/charcoal/conf/env-sourmash.yml +++ b/charcoal/conf/env-sourmash.yml @@ -4,11 +4,11 @@ channels: - bioconda - defaults dependencies: - - python>=3.6 + - python>=3.7,<3.10 - pytest>6,<7 - pytest-dependency>=0.5.1 - pyyaml>5.3,<6 - - sourmash>=4.2.0,<5 + - sourmash>=4.2.2,<5 - pip - pip: - git+https://github.com/dib-lab/charcoal.git diff --git a/charcoal/contigs_list_contaminants.py b/charcoal/contigs_list_contaminants.py index 416985e..d3c9fad 100644 --- a/charcoal/contigs_list_contaminants.py +++ b/charcoal/contigs_list_contaminants.py @@ -82,12 +82,20 @@ def main(args): # load the genome signature genome_sig = sourmash.load_one_signature(args.genome_sig) - # load all of the matches from search --containment in the database - try: - siglist = list(sourmash.load_file_as_signatures(args.matches_sig)) - except ValueError: - siglist = [] - print(f"loaded {len(siglist)} matches from '{args.matches_sig}'") + # load the matches from prefetch as a picklist + picklist = sourmash.picklist.SignaturePicklist('prefetch') + picklist.load(args.matches_csv, picklist.column_name) + + # load all of the matches in the database, as found by prefetch; + # select on them; and then aggregate into MultiIndex. + # CTB note: currently, this loads all the signatures into memory. + # Alternatively we could do something with LazyLoadedIndex maybe? + + siglist = [] + for filename in args.databases: + db = sourmash.load_file_as_index(filename) + db = db.select(picklist=picklist) + siglist += list(db.signatures()) # Hack for examining members of our search database: remove exact matches. new_siglist = [] @@ -192,7 +200,9 @@ def cmdline(sys_args): p = argparse.ArgumentParser(sys_args) p.add_argument('--genome', help='genome file', required=True) p.add_argument('--genome-sig', help='genome sig', required=True) - p.add_argument('--matches-sig', help='all relevant matches', required=True) + p.add_argument('--matches-csv', help='all relevant matches', required=True) + p.add_argument('--databases', help='sourmash databases', required=True, + nargs='+') p.add_argument('--lineages-csv', help='lineage spreadsheet', required=True) p.add_argument('--hitlist', help='hitlist spreadsheet', required=True) p.add_argument('--force', help='continue past survivable errors', diff --git a/charcoal/contigs_search_taxonomy.py b/charcoal/contigs_search_taxonomy.py index 9d83053..239913e 100644 --- a/charcoal/contigs_search_taxonomy.py +++ b/charcoal/contigs_search_taxonomy.py @@ -31,12 +31,22 @@ def main(args): # load the genome signature genome_sig = sourmash.load_one_signature(args.genome_sig) - # load all of the matches from search --containment in the database - try: - siglist = list(sourmash.load_file_as_signatures(args.matches_sig)) - except (ValueError, AssertionError) as e: - siglist = [] - print(f"loaded {len(siglist)} matches from '{args.matches_sig}'") + # load the matches from prefetch as a picklist + picklist = sourmash.picklist.SignaturePicklist('prefetch') + picklist.load(args.matches_csv, picklist.column_name) + + # load all of the matches in the database, as found by prefetch; + # select on them; and then aggregate into MultiIndex. + # CTB note: currently, this loads all the signatures into memory. + # Alternatively we could do something with LazyLoadedIndex maybe? + + siglist = [] + for filename in args.databases: + db = sourmash.load_file_as_index(filename) + db = db.select(picklist=picklist) + siglist += list(db.signatures()) + + print(f"loaded {len(siglist)} matches from '{args.matches_csv}'") # Hack for examining members of our search database: remove exact matches. new_siglist = [] @@ -120,7 +130,9 @@ def cmdline(sys_args): p = argparse.ArgumentParser(sys_args) p.add_argument('--genome', help='genome file', required=True) p.add_argument('--genome-sig', help='genome sig', required=True) - p.add_argument('--matches-sig', help='all relevant matches', required=True) + p.add_argument('--matches-csv', help='all relevant matches', required=True) + p.add_argument('--databases', help='sourmash databases', required=True, + nargs='+') p.add_argument('--lineages-csv', help='lineage spreadsheet', required=True) p.add_argument('--force', help='continue past survivable errors', action='store_true') diff --git a/environment.yml b/environment.yml index 3223f23..38b0b4a 100644 --- a/environment.yml +++ b/environment.yml @@ -4,10 +4,10 @@ channels: - bioconda - defaults dependencies: - - python>=3.7 + - python>=3.7,<3.10 - snakemake-minimal=6.5.1 - screed - click>=7,<8 - pip - mamba - - sourmash>=4.2.0,<5 + - sourmash>=4.2.2,<5 diff --git a/tests/test-data/2.fa.gz.gather-matches.csv b/tests/test-data/2.fa.gz.gather-matches.csv new file mode 100644 index 0000000..7992a54 --- /dev/null +++ b/tests/test-data/2.fa.gz.gather-matches.csv @@ -0,0 +1,2 @@ +intersect_bp,jaccard,max_containment,f_query_match,f_match_query,match_filename,match_name,match_md5,match_bp,query_filename,query_name,query_md5,query_bp +2740000,1.0,1.0,1.0,1.0,,"CP001071.1 Akkermansia muciniphila ATCC BAA-835, complete genome",318839c2,2740000,tests/test-data/genomes/2.fa.gz,"CP001071.1 Akkermansia muciniphila ATCC BAA-835, complete genome",318839c2,2740000 diff --git a/tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.csv b/tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.csv new file mode 100644 index 0000000..497ec72 --- /dev/null +++ b/tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.csv @@ -0,0 +1,19 @@ 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a/tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.sig b/tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.sig deleted file mode 100644 index 0d6bd5d..0000000 --- a/tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.sig +++ /dev/null @@ -1 +0,0 @@ -[{"class":"sourmash_signature","email":"","hash_function":"0.murmur64","filename":"/home/ctbrown/gtdbtk/release89/fastani/database/GCF_002159845.1_genomic.fna.gz","name":"GCF_002159845 s__Anaeromassilibacillus 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\ No newline at end of file diff --git a/tests/test_compare_taxonomy.py b/tests/test_compare_taxonomy.py index 34a13e7..78be56b 100644 --- a/tests/test_compare_taxonomy.py +++ b/tests/test_compare_taxonomy.py @@ -22,6 +22,7 @@ def test_basic(location): args.hit_list = hitlist args.genome_summary = summary_csv args.lineages_csv = utils.relative_file("tests/test-data/test-match-lineages.csv") + args.databases = [utils.relative_file('tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.zip')] args.provided_lineages = None args.contam_summary_json = contam_json args.min_f_ident = compare_taxonomy.F_IDENT_THRESHOLD @@ -30,7 +31,7 @@ def test_basic(location): args.genome = 'loomba' shutil.copyfile(utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.sig"), os.path.join(location, "loomba.sig")) - shutil.copyfile(utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.zip"), os.path.join(location, "loomba.matches.zip")) + shutil.copyfile(utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.csv"), os.path.join(location, "loomba.matches.csv")) shutil.copyfile(utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.contigs-tax.json"), os.path.join(location, "loomba.contigs-tax.json")) status = compare_taxonomy.main(args) diff --git a/tests/test_contigs_list_contaminants.py b/tests/test_contigs_list_contaminants.py index 89130a0..1af4775 100644 --- a/tests/test_contigs_list_contaminants.py +++ b/tests/test_contigs_list_contaminants.py @@ -4,13 +4,38 @@ from charcoal import contigs_list_contaminants + @utils.in_tempdir def test_1_loomba(location): # regression test/check for same results on Loomba args = utils.Args() args.genome = utils.relative_file("demo/genomes/LoombaR_2017__SID1050_bax__bin.11.fa.gz") args.genome_sig = utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.sig") - args.matches_sig = utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.abund.zip") + args.matches_csv = utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.csv") + args.databases = [utils.relative_file('tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.zip')] + args.lineages_csv = utils.relative_file("tests/test-data/test-match-lineages.csv") + args.hitlist = utils.relative_file("tests/test-data/loomba-hit-list.csv") + args.json_out = os.path.join(location, 'tax.json') + args.match_rank = 'genus' + + status = contigs_list_contaminants.main(args) + + assert status == 0 + assert os.path.exists(args.json_out) + + with open(args.json_out, 'rt') as fp: + results = json.load(fp) + assert results != {} + + +@utils.in_tempdir +def test_1_loomba_abund(location): + # regression test/check for same results on Loomba - with abund sigs + args = utils.Args() + args.genome = utils.relative_file("demo/genomes/LoombaR_2017__SID1050_bax__bin.11.fa.gz") + args.genome_sig = utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.sig") + args.matches_csv = utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.csv") + args.databases = [utils.relative_file('tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.abund.zip')] args.lineages_csv = utils.relative_file("tests/test-data/test-match-lineages.csv") args.hitlist = utils.relative_file("tests/test-data/loomba-hit-list.csv") args.json_out = os.path.join(location, 'tax.json') diff --git a/tests/test_contigs_search.py b/tests/test_contigs_search.py index 143b4b5..3f74008 100644 --- a/tests/test_contigs_search.py +++ b/tests/test_contigs_search.py @@ -11,7 +11,8 @@ def test_1(location): args = utils.Args() args.genome = utils.relative_file("tests/test-data/genomes/2.fa.gz") args.genome_sig = utils.relative_file("tests/test-data/genomes/2.fa.gz.sig") - args.matches_sig = utils.relative_file("tests/test-data/2.fa.gz.gather-matches.zip") + args.matches_csv = utils.relative_file("tests/test-data/2.fa.gz.gather-matches.csv") + args.databases = [ utils.relative_file("tests/test-data/2.fa.gz.gather-matches.zip") ] args.lineages_csv = utils.relative_file("tests/test-data/test-match-lineages.csv") args.json_out = os.path.join(location, 'tax.json') args.match_rank = 'genus' @@ -32,7 +33,8 @@ def test_2_loomba(location): args = utils.Args() args.genome = utils.relative_file("demo/genomes/LoombaR_2017__SID1050_bax__bin.11.fa.gz") args.genome_sig = utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.sig") - args.matches_sig = utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.zip") + args.matches_csv = utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.csv") + args.databases = [ utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.zip") ] args.lineages_csv = utils.relative_file("tests/test-data/test-match-lineages.csv") args.json_out = os.path.join(location, 'tax.json') args.match_rank = 'genus' @@ -57,7 +59,8 @@ def test_3_loomba(location): args = utils.Args() args.genome = utils.relative_file("demo/genomes/LoombaR_2017__SID1050_bax__bin.11.fa.gz") args.genome_sig = utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.sig") - args.matches_sig = utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.abund.zip") + args.matches_csv = utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.csv") + args.databases = [ utils.relative_file("tests/test-data/loomba/LoombaR_2017__SID1050_bax__bin.11.fa.gz.matches.zip") ] args.lineages_csv = utils.relative_file("tests/test-data/test-match-lineages.csv") args.json_out = os.path.join(location, 'tax.json') args.match_rank = 'genus' diff --git a/tests/test_snakemake.py b/tests/test_snakemake.py index 48eb8e3..52a43d0 100644 --- a/tests/test_snakemake.py +++ b/tests/test_snakemake.py @@ -71,7 +71,7 @@ def test_make_sig(genome_file): @pytest.mark.parametrize("genome_file", demo_genomes) def test_make_gather_matches(request, genome_file): depends(request, [f"test_make_sig[{g}]" for g in demo_genomes]) - target = f'stage1/{genome_file}.matches.zip' + target = f'stage1/{genome_file}.matches.csv' status, out, err = _run_snakemake_test('demo/demo.conf', target) assert status == 0