def excise_precursors():
global file_genome, parsed_arf, dir_tmp, stack_height_min, dir_tmp, max_pres
# excise precursors from the genome
pprint("#excising precursors\n")
print_stderr("#excising precursors\n")
start()
ret_excise_precursors = None
if options.get('-a'):
cmd = "excise_precursors.py {} {}/{}_parsed.arf {}/precursors.coords -a {} > {}/precursors.fa\n\n".format(
file_genome, dir_tmp, parsed_arf, dir_tmp, stack_height_min,
dir_tmp)
print_stderr(cmd)
ret_excise_precursors = os.popen(cmd).read()
else:
cmd = "excise_precursors_iterative_final.py {} {}/{}_parsed.arf {}/precursors.fa {}/precursors.coords {}\n".format(
file_genome, dir_tmp, parsed_arf, dir_tmp, dir_tmp, max_pres)
print_stderr(cmd)
ret_excise_precursors = os.popen(cmd).read()
fname = '{}/precursors.fa_stack'.format(dir_tmp)
OSS = open_or_die2(fname, 'rb')
stack_height_min = OSS.readline().strip()
OSS.close()
end()
fname = '{}/precursors.fa'.format(dir_tmp)
# if (-z "$dir_tmp/precursors.fa" or not -f "$dir_tmp/precursors.fa"):
if not file_s(fname) or not os.path.isfile(
fname): # empty or not a regular plain file
die("No precursors excised\n")
return 0
def parse_file_fasta_seqkey(file_fasta, hsh, options):
if options.get('-a') == '':
print_stderr('reading file into hash\n')
_id = ''
seq = ''
running_1 = 0
FASTA = open_or_die2(file_fasta, 'rb')
while True:
l = FASTA.readline().strip()
if not l:
break
m = re.match(r'^>(\S+)', l)
if m:
_id = m.group()
seq = ''
while True:
ll = FASTA.readline().strip()
if not ll:
break
mm = re.match(r'^>(\S+)', ll)
if mm:
cnt = find_cnt(_id)
seq = tr(seq, '[acgtun.]', '[ACGTTNN]')
# ATTR: Performance issue below:
# create_hash_key_chain(hsh, 0, seq)
try:
hsh[seq] = (hsh[seq]) + cnt
except KeyError:
hsh[seq] = cnt
running_1 += 1
if options.get('-a') == '':
print_stderr('{}\r'.format(running_1))
_id = mm.group()
seq = ''
continue
seq += ll
cnt = find_cnt(_id)
seq = tr(seq, '[acgtun.]', '[ACGTTNN]')
create_hash_key_chain(hsh, 0, seq)
hsh[seq] += cnt
running_1 += 1
if options.get('-a') == '':
print_stderr('{}\r'.format(running_1))
FASTA.close()
if re.search(r'\s+', _id):
die('Error in line {}: The identifier\n {}\n\ncontains white spaces\n\n{}\n\nYou could run remove_white_space_in_id.py inputfile > newfile\nThis will remove everything from the id line after the first whitespace\n'
.format(Nicenumber(counter), _id, hint))
else:
create_hash_key_chain(hash_num, 0, _id)
hash_num[_id] += 1
elif not re.match(r'^([A|C|G|T|U|N|a|c|g|t|u|n]+)$', rin):
die('Error in line {}: The sequence\n{}\n\ncontains characters others than [acgtunACGTUN]\n\n{}'
.format(Nicenumber(counter), rin, hint))
if __name__ == '__main__':
hash_num = {}
_id = None
hint = 'Please check your file for the following issues:\n\nI. Sequences are allowed only to comprise characters [ACGTNacgtn].\nII. Identifiers are not allowed to have withespaces.\n'
if len(sys.argv) == 1:
# from stdin
read_handler(sys.stdin)
else:
# from files
for f in sys.argv[1:]:
IN = open_or_die2(f, 'rb')
read_handler(IN)
IN.close()
sys.exit(0)
line = []
thres = -50
if options.get('-s') is not None:
thres = options.get('-s')
score = thres
_max = 'na'
maxs = 999999999999999999999999999
if options.get('-t'):
_max = options.get('-t')
maxs = _max
IN = open_or_die2(options.get('-r'), 'rb')
seqcol = 15
if options.get('-m') == '':
seqcol = 13
if options.get('-k') == '':
seqcol = 14
names = ('0', '1', '2', '3', '4', '5', '6', '7', '8', '9', '10', '11',
'12', 'mature', 'star', 'pres')
while True:
l = IN.readline()
if not l:
break
def test_input_files():
global file_reads, file_reads_vs_genome, file_genome, file_precursors, minpreslen, file_mature_ref_other_species, file_mature_ref_this_species
IN = open_or_die2(file_reads, 'rb')
line = IN.readline().strip()
if not re.search(r'^>\S+', line):
printErr()
die("The first line of file $file_reads does not start with '>identifier'\nReads file {} is not a valid fasta file\n\n"
.format(file_reads))
if re.search(r'\s', line):
printErr()
die('File {} contains not allowed characters in sequences\nAllowed characters are ACGTUN\nReads file {} is not a fasta file\n\n'
.format(file_reads, file_reads))
line = IN.readline()
if not re.search(r'^[ACGTUNacgtun]*$', line):
printErr()
die('File {} contains not allowed characters in sequences\nAllowed characters are ACGTUN\nReads file {} is not a fasta file\n\n'
.format(file_reads, file_reads))
IN.close()
IN = open_or_die2(file_genome, 'rb')
line = IN.readline().strip()
if not re.search(r'>\S+', line):
printErr()
die("The first line of file {} does not start with '>identifier'\nGenome file {} is not a fasta file\n\n"
.format(file_genome, file_genome))
if re.search(r'\s', line):
printErr()
die('Genome file {} has not allowed whitespaces in its first identifier\n\n'
.format(file_genome))
# get genome ids
tmps = os.popen('grep ">" {}'.format(file_genome)).read().strip()
genomeids = dict(map(lambda x: (x, 1), re.split("\n", tmps)))
line = IN.readline()
if not re.search(r'^[ACGTUNacgtun]*$', line):
printErr()
die('File {} contains not allowed characters in sequences\nAllowed characters are ACGTUN\nGenome file {} is not a fasta file\n\n'
.format(file_genome, file_genome))
IN.close()
IN = open_or_die2(file_reads_vs_genome, 'rb')
line = IN.readline()
if not re.search(
r'^(\S+_x\d+)\s+(\d+)\s+(\d+)\s+(\d+)\s+(\S+)\s+(\S+)\s+(\d+)\s+(\d+)\s+(\d+)\s+(\S+)\s+([+-])\s+(\d+)\s*([mDIM]*)$',
line):
printErr()
die('Mapping file {} is not in arf format\n\nEach line of the mapping file must consist of the following fields\nreadID_wo_whitespaces length start end read_sequence genomicID_wo_whitspaces length start end genomic_sequence strand #mismatches editstring\nThe editstring is optional and must not be contained\nThe readID must end with _xNumber and is not allowed to contain whitespaces.\nThe genomeID is not allowed to contain whitespaces.'
.format(file_reads_vs_genome))
IN.close()
# get ids from arf file and compare them with ids from the genome file
tmps = os.popen(
'cut -f6 {}|sort -u'.format(file_reads_vs_genome)).read().strip()
for s in re.split("\n", tmps):
if not genomeids.get(">{}".format(s)):
die("The mapped reference id {} from file {} is not an id of the genome file {}\n\n"
.format(s, file_reads_vs_genome, file_genome))
if not re.search('none', file_mature_ref_this_species):
IN = open_or_die2(file_mature_ref_this_species, 'rb')
line = IN.readline().strip()
if not re.search(r'>\S+', line):
printErr()
die("The first line of file {} does not start with '>identifier'\nmiRNA reference this species file {} is not a fasta file\n\n"
.format(file_mature_ref_this_species,
file_mature_ref_this_species))
if re.search(r'\s', line):
printErr()
die("miRNA reference this species file {} has not allowed whitespaces in its first identifier\n\n"
.format(file_mature_ref_this_species))
line = IN.readline()
if not re.search(r'^[ACGTUNacgtun]*$', line):
printErr()
die("File {} contains not allowed characters in sequences\nAllowed characters are ACGTUN\nmiRNA reference this species file {} is not a fasta file\n\n"
.format(file_mature_ref_this_species,
file_mature_ref_this_species))
IN.close()
if not re.search('none', file_mature_ref_other_species):
IN = open_or_die2(file_mature_ref_other_species, 'rb')
line = IN.readline().strip()
if not re.search(r'>\S+', line):
printErr()
die("The first line of file {} does not start with '>identifier'\nmiRNA reference this species file {} is not a fasta file\n\n"
.format(file_mature_ref_other_species,
file_mature_ref_other_species))
if re.search(r'\s', line):
printErr()
die("miRNA reference this species file {} has not allowed whitespaces in its first identifier\n\n"
.format(file_mature_ref_other_species))
line = IN.readline()
if not re.search(r'^[ACGTUNacgtun]*$', line):
printErr()
die("File {} contains not allowed characters in sequences\nAllowed characters are ACGTUN\nmiRNA reference this species file {} is not a fasta file\n\n"
.format(file_mature_ref_other_species,
file_mature_ref_other_species))
IN.close()
if not re.search('none', file_precursors):
IN = open_or_die2(file_precursors, 'rb')
line = IN.readline().strip()
if not re.search(r'>\S+', line):
printErr()
die("The first line of file {} does not start with '>identifier'\nmiRNA reference this species file {} is not a fasta file\n\n"
.format(file_precursors, file_precursors))
if re.search(r'\s', line):
printErr()
die("precursor file {} has not allowed whitespaces in its first identifier\n\n"
.format(file_precursors))
line = IN.readline()
if not re.search(r'^[ACGTUNacgtun]*$', line):
printErr()
die("File {} contains not allowed characters in sequences\nAllowed characters are ACGTUN\nmiRNA reference this species file {} is not a fasta file\n\n"
.format(file_precursors, file_precursors))
if len(line) < minpreslen:
printErr()
die("The precursor file {} does not contain sequences of at least {} nt\nPlease make sure that you provided the correct file and the correct parameter ordering when calling {}\nIf you have precursors with less than {} please use option -p <int> to specify this length\n"
.format(file_precursors, minpreslen, sys.argv[0], minpreslen))
IN.close()
# #################################################
# precheck finished
# #################################################
# do stringent testing of all input files
pprint("#testing input files\n")
print_stderr("#testing input files\n")
if not re.search('none', file_mature_ref_this_species):
start()
cmd = "sanity_check_mature_ref.py {} 2>&1\n\n".format(
file_mature_ref_this_species)
print_stderr(cmd)
ret_file_mature_ref_this_species = os.popen(cmd).read().strip()
if ret_file_mature_ref_this_species:
printErr()
die("problem with {} {}\n".format(
file_mature_ref_this_species,
ret_file_mature_ref_this_species))
end()
if not re.search(r'none', file_mature_ref_other_species):
start()
cmd = "sanity_check_mature_ref.py {} 2>&1\n\n".format(
file_mature_ref_other_species)
print_stderr(cmd)
ret_file_mature_ref_other_species = os.popen(cmd).read().strip()
if ret_file_mature_ref_other_species:
printErr()
die("problem with {} {}\n".format(
file_mature_ref_other_species,
ret_file_mature_ref_other_species))
end()
cmd = "sanity_check_reads_ready_file.py {} 2>&1\n\n".format(file_reads)
print_stderr(cmd)
start()
ret_test_file_reads = os.popen(cmd).read().strip()
if ret_test_file_reads:
printErr()
die("problem with {} {}\n".format(file_reads, ret_test_file_reads))
end()
start()
cmd = "sanity_check_genome.py {} 2>&1;\n\n".format(file_genome)
print_stderr(cmd)
ret_test_file_genome = os.popen(cmd).read().strip()
if ret_test_file_genome:
printErr()
die("problem with {} {}\n".format(file_genome, ret_test_file_genome))
end()
start()
cmd = "sanity_check_mapping_file.py {} 2>&1".format(file_reads_vs_genome)
print_stderr(cmd)
ret_test_file_reads_genome = os.popen(cmd).read().strip()
if ret_test_file_reads_genome:
printErr()
die("problem with {} {}\n".format(file_reads_vs_genome,
ret_test_file_reads_genome))
end()
if not re.search('none', file_precursors):
start()
cmd = "sanity_check_mature_ref.py {} 2>&1".format(file_precursors)
print_stderr(cmd)
ret_file_precursors = os.popen(cmd).read().strip()
if ret_file_precursors:
printErr()
die("problem with {} {}\n".format(file_precursors,
ret_file_precursors))
end()
start()
if not re.search('none', file_mature_ref_this_species, re.IGNORECASE):
print_stderr("Quantitation of expressed miRNAs in data\n\n\n")
species = ''
if options.get('-t'):
species = "-t {}".format(options.get('-t'))
file_star = ''
if options.get('-s'):
if file_s(options.get('-s')):
file_star = "-s {}".format(options.get('-s'))
else:
print_stderr(
"File {} specified by option -s is empty or not found\n"
.format(options.get('-s')))
options['-s'] = 0
print("#Quantitation of known miRNAs in data\n")
dopt = ""
Popt = ""
if options.get('-d') == '':
dopt = "-d"
if options.get('-P') == '':
Popt = "-P"
quant = "quantifier.py -p {} -m {} -r {} {} {} -y {} -k {} {}".format(
file_precursors, file_mature_ref_this_species, file_reads,
file_star, species, ltime, dopt, Popt)
print_stderr(quant, "\n")
os.system(quant)
options[
'-q'] = "expression_analyses/expression_analyses_{}/miRBase.mrd".format(
ltime)
end()
else:
print_stderr(
"Pre-quantitation is skipped caused by missing file with known miRNAs\n\n\n"
)
else:
print_stderr(
"Pre-quantitation is skipped caused by missing file with known precursor miRNAs\n\n\n"
)