def combine_rerun_with_orig(self):
"""Special case when a sample with low reads was rerun in an other pool.
Run this just before the combine_reads() method of the associated cluster.
This method is called on the reruned sampled, not the original."""
# Check we have a rerun #
if self.info.get('rerun') is None: return False
# Check we are processed #
assert self.fasta.count > 0
# Get the original sample #
run, pool, num = self.info['rerun']['run'], self.info['rerun']['pool'], self.info['rerun']['num']
orig_sample = illumitag.runs[run][pool-1][num-1]
merged = FASTA(orig_sample.base_dir + 'rerun_merged.fasta')
# Check we don't merge twice #
assert orig_sample.count == orig_sample.fasta.count
# Do it #
merged.create()
merged.add(orig_sample.fasta)
merged.add(self.fasta)
merged.close()
merged.rename_with_num(orig_sample.name + '_read', orig_sample.fasta)
merged.remove()
# Check #
orig_sample.fasta = FASTA(orig_sample.fasta.path)
assert orig_sample.count < orig_sample.fasta.count
return True
def fasta(self):
"""The fasta file containing the filtered genes of this cluster
The names now will correspond to long descriptive names"""
fasta = FASTA(self.p.fasta)
if not fasta:
fasta.create()
for gene in self.filtered_genes: fasta.add_str(str(gene), name=gene.name)
fasta.close()
return fasta
def fasta(self):
"""The fasta file containing the filtered genes of this cluster
The names now will correspond to long descriptive names"""
fasta = FASTA(self.p.fasta)
if not fasta:
fasta.create()
for gene in self.filtered_genes:
fasta.add_str(str(gene), name=gene.name)
fasta.close()
return fasta
def fasta(self):
"""Make a fasta file with all uniprot proteins that are related to
this family."""
fasta = FASTA(self.p.proteins)
if not fasta.exists:
fasta.create()
for seq in pfam.fasta:
if self.fam_name in seq.description: fasta.add_seq(seq)
fasta.close()
assert fasta
# Return #
return fasta
def fasta(self):
"""Make a fasta file with all uniprot proteins that are related to
this family."""
fasta = FASTA(self.p.proteins)
if not fasta.exists:
fasta.create()
for seq in pfam.fasta:
if self.fam_name in seq.description: fasta.add_seq(seq)
fasta.close()
assert fasta
# Return #
return fasta
def test(self):
"""Search one sequence, and see if it works."""
# New directory #
directory = new_temp_dir()
# A randomly chosen sequence (H**o sapiens mRNA for prepro cortistatin) #
seq = """ACAAGATGCCATTGTCCCCCGGCCTCCTGCTGCTGCTGCTCTCCGGGGCCACGGCCACCGCTGCCCTGCC
CCTGGAGGGTGGCCCCACCGGCCGAGACAGCGAGCATATGCAGGAAGCGGCAGGAATAAGGAAAAGCAGC
CTCCTGACTTTCCTCGCTTGGTGGTTTGAGTGGACCTCCCAGGCCAGTGCCGGGCCCCTCATAGGAGAGG
AAGCTCGGGAGGTGGCCAGGCGGCAGGAAGGCGCACCCCCCCAGCAATCCGCGCGCCGGGACAGAATGCC
CTGCAGGAACTTCTTCTGGAAGACCTTCTCCTCCTGCAAATAAAACCTCACCCATGAATGCTCACGCAAG
TTTAATTACAGACCTGAA"""
seq = seq.replace('\n','')
seq = seq.replace(' ','')
# Make input #
input_fasta = FASTA(directory + 'input.fasta')
input_fasta.create()
input_fasta.add_str(seq, "My test sequence")
input_fasta.close()
# Make output #
out_path = directory + 'output.blast'
# Make extras parameters #
params = {'-outfmt': 0,
'-evalue': 1e-5,
'-perc_identity': 99}
# Make the search #
search = SeqSearch(input_fasta,
self.blast_db,
'nucl',
'blast',
num_threads = 1,
out_path = out_path,
params = params)
# Run it #
search.run()
# Print result #
print "Success", directory
def test(self):
"""Search one sequence, and see if it works."""
# New directory #
directory = new_temp_dir()
# A randomly chosen sequence (H**o sapiens mRNA for prepro cortistatin) #
seq = """ACAAGATGCCATTGTCCCCCGGCCTCCTGCTGCTGCTGCTCTCCGGGGCCACGGCCACCGCTGCCCTGCC
CCTGGAGGGTGGCCCCACCGGCCGAGACAGCGAGCATATGCAGGAAGCGGCAGGAATAAGGAAAAGCAGC
CTCCTGACTTTCCTCGCTTGGTGGTTTGAGTGGACCTCCCAGGCCAGTGCCGGGCCCCTCATAGGAGAGG
AAGCTCGGGAGGTGGCCAGGCGGCAGGAAGGCGCACCCCCCCAGCAATCCGCGCGCCGGGACAGAATGCC
CTGCAGGAACTTCTTCTGGAAGACCTTCTCCTCCTGCAAATAAAACCTCACCCATGAATGCTCACGCAAG
TTTAATTACAGACCTGAA"""
seq = seq.replace('\n','')
seq = seq.replace(' ','')
# Make input #
input_fasta = FASTA(directory + 'input.fasta')
input_fasta.create()
input_fasta.add_str(seq, "My test sequence")
input_fasta.close()
# Make output #
out_path = directory + 'output.blast'
# Make extras parameters #
params = {'-outfmt': 0,
'-evalue': 1e-5,
'-perc_identity': 99}
# Make the search #
search = SeqSearch(input_fasta,
self.blast_db,
'nucl',
'blast',
num_threads = 1,
out_path = out_path,
params = params)
# Run it #
search.run()
# Print result #
print("Success", directory)
fnas_genes = [strip(seq) for seq in fna]
print faa, len(set(fnas_genes) ^ set(faas_genes)), "discrepancies"
#print "- in fna but not in faa:", [x for x in set(fnas_genes) - set(faas_genes)]
#print "- in faa but not in fna:", [x for x in set(faas_genes) - set(fnas_genes)]
#print ""
fnas_genes = [strip(seq) for fna in fnas for seq in fna]
print len(fnas_genes), len(set(fnas_genes))
for genome in faas:
out_path = genomes_dir + genome.short_prefix + '.fasta'
out_fasta = FASTA(out_path)
out_fasta.create()
for seq in genome:
out_fasta.add_str(str(seq.seq), strip(seq))
out_fasta.close()
out_fasta.gzip_to()
out_fasta.remove()
def lines():
for genome in faas:
for gene in genome:
name = strip(gene)
yield name + '\t' + gene.description[len(name):].rstrip(
' |') + '\n'
annotations_path = current_dir + '../ld12/data/annotations.tsv'
with open(annotations_path, 'w') as handle:
handle.writelines(lines())
class PairedFASTA(object):
"""Read and write FASTA file pairs without using too much RAM"""
format = 'fasta'
def __len__(self):
return self.count
def __iter__(self):
return self.parse()
def __nonzero__(self):
return bool(self.fwd) and bool(self.rev)
def __repr__(self): return '<%s object on "%s" and "%s">' % \
(self.__class__.__name__, self.fwd.path, self.rev.path)
def __enter__(self):
return self.create()
def __exit__(self, exc_type, exc_value, traceback):
self.close()
@property
def exists(self):
return self.fwd.exists and self.rev.exists
def __init__(self, fwd, rev, parent=None):
# FASTA objects #
self.fwd = FASTA(fwd)
self.rev = FASTA(rev)
# Extra #
self.gzipped = self.fwd.gzipped
self.parent = parent
@property_cached
def count(self):
assert self.fwd.count == self.rev.count
return self.fwd.count
def open(self):
self.fwd.open()
self.rev.open()
def parse(self):
return izip(self.fwd.parse(), self.rev.parse())
def close(self):
self.fwd.close()
self.rev.close()
def create(self):
self.fwd.create()
self.rev.create()
return self
def add(self, f, r):
return self.add_pair((f, r))
def add_pair(self, pair):
self.fwd.add_seq(pair[0])
self.rev.add_seq(pair[1])
def remove(self):
self.fwd.remove()
self.rev.remove()
@property
def progress(self):
"""Just like self.parse but display a progress bar"""
return tqdm(self, total=len(self))
def subsample(self, down_to, dest_pair=None):
# Check size #
assert down_to < len(self)
# Make new pair of files #
if dest_pair is None:
dest_fwd_path = self.fwd_path.new_name_insert("subsampled")
dest_rev_path = self.rev_path.new_name_insert("subsampled")
dest_pair = self.__class__(dest_fwd_path, dest_rev_path)
# Do it #
dest_pair.create()
for pair in isubsample(self, down_to):
dest_pair.add_pair(pair)
self.subsampled.close()
# Did it work #
assert len(dest_pair) == down_to
#------------------------------- Extensions ------------------------------#
def parse_primers(self, *args, **kwargs):
fwd_gen = self.fwd.parse_primers(*args, **kwargs)
rev_gen = self.rev.parse_primers(*args, **kwargs)
generator = izip(fwd_gen, rev_gen)
return GenWithLength(generator, len(fwd_gen))
class PairedFASTA(object):
"""Read and write FASTA file pairs without using too much RAM"""
format = 'fasta'
def __len__(self): return self.count
def __iter__(self): return self.parse()
def __nonzero__(self): return bool(self.fwd) and bool(self.rev)
def __repr__(self): return '<%s object on "%s" and "%s">' % \
(self.__class__.__name__, self.fwd.path, self.rev.path)
def __enter__(self): return self.create()
def __exit__(self, exc_type, exc_value, traceback): self.close()
@property
def exists(self): return self.fwd.exists and self.rev.exists
def __init__(self, fwd, rev, parent=None):
# FASTA objects #
self.fwd = FASTA(fwd)
self.rev = FASTA(rev)
# Extra #
self.gzipped = self.fwd.gzipped
self.parent = parent
@property_cached
def count(self):
assert self.fwd.count == self.rev.count
return self.fwd.count
def open(self):
self.fwd.open()
self.rev.open()
def parse(self):
return izip(self.fwd.parse(), self.rev.parse())
def close(self):
self.fwd.close()
self.rev.close()
def create(self):
self.fwd.create()
self.rev.create()
return self
def add(self, f, r):
return self.add_pair((f,r))
def add_pair(self, pair):
self.fwd.add_seq(pair[0])
self.rev.add_seq(pair[1])
def remove(self):
self.fwd.remove()
self.rev.remove()
@property
def progress(self):
"""Just like self.parse but display a progress bar"""
return tqdm(self, total=len(self))
def subsample(self, down_to, dest_pair=None):
# Check size #
assert down_to < len(self)
# Make new pair of files #
if dest_pair is None:
dest_fwd_path = self.fwd_path.new_name_insert("subsampled")
dest_rev_path = self.rev_path.new_name_insert("subsampled")
dest_pair = self.__class__(dest_fwd_path, dest_rev_path)
# Do it #
dest_pair.create()
for pair in isubsample(self, down_to): dest_pair.add_pair(pair)
self.subsampled.close()
# Did it work #
assert len(dest_pair) == down_to
#------------------------------- Extensions ------------------------------#
def parse_primers(self, *args, **kwargs):
fwd_gen = self.fwd.parse_primers(*args, **kwargs)
rev_gen = self.rev.parse_primers(*args, **kwargs)
generator = izip(fwd_gen, rev_gen)
return GenWithLength(generator, len(fwd_gen))
seq = seq.split('[')[0]
return seq
for faa,fna in zip(faas, fnas):
faas_genes = [strip(seq) for seq in faa]
fnas_genes = [strip(seq) for seq in fna]
print faa, len(set(fnas_genes) ^ set(faas_genes)), "discrepancies"
#print "- in fna but not in faa:", [x for x in set(fnas_genes) - set(faas_genes)]
#print "- in faa but not in fna:", [x for x in set(faas_genes) - set(fnas_genes)]
#print ""
fnas_genes = [strip(seq) for fna in fnas for seq in fna]
print len(fnas_genes), len(set(fnas_genes))
for genome in faas:
out_path = genomes_dir + genome.short_prefix + '.fasta'
out_fasta = FASTA(out_path)
out_fasta.create()
for seq in genome: out_fasta.add_str(str(seq.seq), strip(seq))
out_fasta.close()
out_fasta.gzip_to()
out_fasta.remove()
def lines():
for genome in faas:
for gene in genome:
name = strip(gene)
yield name + '\t' + gene.description[len(name):].rstrip(' |') + '\n'
annotations_path = current_dir + '../ld12/data/annotations.tsv'
with open(annotations_path, 'w') as handle: handle.writelines(lines())
class Foraminifera(Database):
"""This is a custom database containing exlcusively Foraminifera sequences.
https://genev.unige.ch/research/laboratory/Jan-Pawlowski
You should place the file "foram_db_cor.fasta" in: ~/databases/foraminifera/
Then you can run this:
from seqsearch.databases.foraminifera import foraminifera
foraminifera.process()
print foraminifera.tax_depth_freq
"""
short_name = "foraminifera"
long_name = 'The custom made Foraminifera database as received by email on 7th April 2017'
all_paths = """
/foram_db_cor.fasta
/foram_mothur.fasta
/foram_mothur.tax
"""
@property
def rank_names(self):
"""The names of the ranks. Total 9 ranks."""
return ['Domain', # 0
'Kingdom', # 1
'Phylum', # 2
'Class', # 3
'Order', # 4
'Family', # 5
'Tribe', # 6
'Genus', # 7
'Species'] # 8
def __init__(self, base_dir=None):
# Base directory #
if base_dir is None: base_dir = home
self.base_dir = base_dir + 'databases/' + self.short_name + '/'
self.p = AutoPaths(self.base_dir, self.all_paths)
# The results #
self.alignment = FASTA(self.p.mothur_fasta)
self.taxonomy = FilePath(self.p.mothur_tax)
# The part that mothur will use for naming files #
self.nickname = "foram_mothur"
def process(self):
# The file that was received by email without documentation T_T #
raw = FASTA(self.p.cor)
# Open files #
self.alignment.create()
self.taxonomy.create()
# Loop #
for seq in raw:
# Parse #
name = seq.id[11:].split('|')
num = name.pop(0)
# Check #
for x in name: assert ';' not in x
for x in name: assert '\t' not in x
# Make ranks #
ranks = ['Eukaryota' , # 0 Domain
'Rhizaria' , # 1 Kingdom
'Foraminifera' , # 2 Phylum
name[0] , # 3 Class
name[1] , # 4 Order
name[2] , # 5 Family
name[3] , # 6 Tribe
name[4] , # 7 Genus
name[5]] # 8 Species
# The taxonomy string #
tax_line = ';'.join(ranks)
# Add sequence to the new fasta file #
self.alignment.add_str(str(seq.seq), name="foram" + num)
# Add the taxonomy to the tax file #
self.taxonomy.add_str("foram" + num + '\t' + tax_line + '\n')
# Close files #
self.alignment.close()
self.taxonomy.close()
class Cluster(object):
"""Analyzes a group of samples."""
all_paths = """
/reads/all_reads.fasta
/otus/
/logs/
/report/report.pdf
/metadata.csv
"""
def __repr__(self): return '<%s object "%s" with %i samples>' % (self.__class__.__name__, self.name, len(self.samples))
def __iter__(self): return iter(self.samples)
def __len__(self): return len(self.samples)
def __getitem__(self, key):
if isinstance(key, basestring): return [c for c in self.children if c.short_name == key.lower()][0]
elif isinstance(key, int) and hasattr(self.first, 'num'): return [c for c in self.children if c.num == key][0]
else: return self.children[key]
@property
def first(self): return self.children[0]
@property
def count_seq(self):
return sum([len(sample) for sample in self])
def __init__(self, samples, name, base_dir=None):
# Save samples #
self.name = name
self.samples, self.children = samples, samples
# Check names are unique #
names = [s.short_name for s in samples if s.used]
assert len(names) == len(set(names))
# Figure out pools #
self.pools = list(set([s.pool for s in self.samples]))
self.pools.sort(key = lambda x: x.id_name)
# Directory #
if base_dir: self.base_dir = base_dir
else: self.base_dir = illumitag.view_dir + "clusters/" + self.name + '/'
# Loaded #
self.loaded = False
def load(self):
"""A second __init__ that is delayed and called only if needed"""
# Load the pools and samples #
for p in self.pools: p.load()
for s in self.samples: s.load()
# Dir #
self.p = AutoPaths(self.base_dir, self.all_paths)
# Figure out if it's a project #
if set(self.samples) == set(self.first.pool.project.samples): self.project = self.first.pool.project
else: self.project = None
# Runner #
self.runner = ClusterRunner(self)
# FASTA #
self.reads = FASTA(self.p.all_reads_fasta)
# OTU picking #
self.otu_uparse = UparseOTUs(self)
self.otu_uclust = UclustOTUs(self)
self.otu_cdhit = CdhitOTUs(self)
# Preferred #
self.otus = self.otu_uparse
# Simple reporting #
self.reporter = ClusterReporter(self)
# Full report #
self.report = ClusterReport(self)
# Loaded #
self.loaded = True
# Return self for convenience #
return self
def run(self, *args, **kwargs):
self.runner.run(*args, **kwargs)
def run_slurm(self, *args, **kwargs):
self.runner.run_slurm(*args, **kwargs)
def process_samples(self):
for sample in tqdm(self): sample.process()
def combine_reads(self):
"""This is the first function should call. It will combine all the
reads of all the samples of this cluster into one big FASTA file."""
paths = [sample.fasta.path for sample in self]
shell_output('cat %s > %s' % (' '.join(paths), self.reads))
return self.reads
def set_size(self, length):
"""Trim all sequences to a specific length starting from the end."""
self.size_trimmed = FASTA(new_temp_path())
def trim_iterator(reads):
for read in reads:
if len(read) < length: continue
yield read[-length:]
self.size_trimmed.write(trim_iterator(self.reads))
self.size_trimmed.close()
# Replace it #
self.reads.remove()
shutil.move(self.size_trimmed, self.reads)
def run_uparse(self): self.otu_uparse.run()
@property
def metadata(self):
return pandas.DataFrame([s.info for s in self], index=[s.short_name for s in self])
def export_metadata(self):
self.metadata.to_csv(self.p.metadata, sep='\t', encoding='utf-8')
# Get clustering values #
r1, r2 = list(set([p.run for p in proj]))
r1.parse_report_xml()
r2.parse_report_xml()
print float(r1.report_stats['fwd']['DensityPF']) / float(r1.report_stats['fwd']['DensityRaw'])
print float(r2.report_stats['fwd']['DensityPF']) / float(r2.report_stats['fwd']['DensityRaw'])
# Check below 400 bp sequences #
folder = DirectoryPath(illumitag.projects['evaluation'].base_dir + "below_400/")
over = FASTA(folder + "reads.fasta")
def over_iterator(reads, max_length=400):
for read in reads:
if len(read) <= max_length: yield read
over.create()
for pool in pools: over.add_iterator(over_iterator(pool.good_barcodes.assembled.good_primers.qual_filtered))
over.close()
over.graphs[-1].plot()
crest = SimpleCrestTaxonomy(over, folder)
crest.assign()
crest.composition.graph.plot()
rdp = SimpleRdpTaxonomy(over, folder)
rdp.assign()
rdp.composition.graph.plot()
# Check unassembled mate pairs #
unassembled = [p.good_barcodes.unassembled for p in pools]
paths = [u.flipped_reads.path for u in unassembled]
folder = DirectoryPath(illumitag.projects['evaluation'].base_dir + "unassembled_taxonomy/")
all_unassembled = FASTA(folder + 'unassembled_reads.fasta')
shell_output('cat %s > %s' % (' '.join(paths), all_unassembled))
tax = SimpleRdpTaxonomy(all_unassembled, folder)
