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#! /usr/bin/env python
# -*- coding: utf-8 -*-
"""
Readqc report: record stat key-value in readqc-stats.txt
### JGI_Analysis_Utility_Illumina::illumina_read_level_report
Created: Jul 24 2013
sulsj (ssul@lbl.gov)
"""
import os
import sys
## custom libs in "../lib/"
srcDir = os.path.dirname(__file__)
sys.path.append(os.path.join(srcDir, 'tools')) ## ./tools
sys.path.append(os.path.join(srcDir, '../lib')) ## rqc-pipeline/lib
sys.path.append(os.path.join(srcDir, '../tools')) ## rqc-pipeline/tools
from readqc_constants import RQCReadQcConfig, ReadqcStats
from rqc_constants import RQCExitCodes
from os_utility import run_sh_command
from common import append_rqc_stats, append_rqc_file
statsFile = RQCReadQcConfig.CFG["stats_file"]
filesFile = RQCReadQcConfig.CFG["files_file"]
"""
Title : read_megablast_hits
Function : This function generates tophit list of megablast against different databases.
Usage : read_megablast_hits(db_name, log)
Args : blast db name or full path
Returns : SUCCESS
FAILURE
Comments :
"""
def read_megablast_hits(db, log):
currentDir = RQCReadQcConfig.CFG["output_path"]
megablastDir = "megablast"
megablastPath = os.path.join(currentDir, megablastDir)
statsFile = RQCReadQcConfig.CFG["stats_file"]
filesFile = RQCReadQcConfig.CFG["files_file"]
##
## Process blast output files
##
matchings = 0
hitCount = 0
parsedFile = os.path.join(megablastPath, "megablast.*.%s*.parsed" % (db))
matchings, _, exitCode = run_sh_command("grep -v '^#' %s 2>/dev/null | wc -l " % (parsedFile), True, log)
if exitCode == 0: ## if parsed file found.
t = matchings.split()
if len(t) == 1 and t[0].isdigit():
hitCount = int(t[0])
append_rqc_stats(statsFile, ReadqcStats.ILLUMINA_READ_MATCHING_HITS + " " + db, hitCount, log)
##
## add .parsed file
##
parsedFileFound, _, exitCode = run_sh_command("ls %s" % (parsedFile), True, log)
if parsedFileFound:
parsedFileFound = parsedFileFound.strip()
append_rqc_file(filesFile, ReadqcStats.ILLUMINA_READ_PARSED_FILE + " " + db, os.path.join(megablastPath, parsedFileFound), log)
else:
log.error("- Failed to add megablast parsed file of %s." % (db))
return RQCExitCodes.JGI_FAILURE
##
## wc the top hits
##
topHit = 0
tophitFile = os.path.join(megablastPath, "megablast.*.%s*.parsed.tophit" % (db))
tophits, _, exitCode = run_sh_command("grep -v '^#' %s 2>/dev/null | wc -l " % (tophitFile), True, log)
t = tophits.split()
if len(t) == 1 and t[0].isdigit():
topHit = int(t[0])
append_rqc_stats(statsFile, ReadqcStats.ILLUMINA_READ_TOP_HITS + " " + db, topHit, log)
##
## wc the taxonomic species
##
spe = 0
taxlistFile = os.path.join(megablastPath, "megablast.*.%s*.parsed.taxlist" % (db))
species, _, exitCode = run_sh_command("grep -v '^#' %s 2>/dev/null | wc -l " % (taxlistFile), True, log)
t = species.split()
if len(t) == 1 and t[0].isdigit():
spe = int(t[0])
append_rqc_stats(statsFile, ReadqcStats.ILLUMINA_READ_TAX_SPECIES + " " + db, spe, log)
##
## wc the top 100 hit
##
top100hits = 0
top100hitFile = os.path.join(megablastPath, "megablast.*.%s*.parsed.top100hit" % (db))
species, _, exitCode = run_sh_command("grep -v '^#' %s 2>/dev/null | wc -l " % (top100hitFile), True, log)
t = species.split()
if len(t) == 1 and t[0].isdigit():
top100hits = int(t[0])
append_rqc_stats(statsFile, ReadqcStats.ILLUMINA_READ_TOP_100HITS + " " + db, top100hits, log)
##
## Find and add taxlist file
##
taxListFound, _, exitCode = run_sh_command("ls %s" % (taxlistFile), True, log)
taxListFound = taxListFound.strip()
if taxListFound:
append_rqc_file(filesFile, ReadqcStats.ILLUMINA_READ_TAXLIST_FILE + " " + db, os.path.join(megablastPath, taxListFound), log)
else:
log.error("- Failed to add megablast taxlist file of %s." % (db))
return RQCExitCodes.JGI_FAILURE
##
## Find and add tophit file
##
tophitFound, _, exitCode = run_sh_command("ls %s" % (tophitFile), True, log)
tophitFound = tophitFound.strip()
if tophitFound:
append_rqc_file(filesFile, ReadqcStats.ILLUMINA_READ_TOPHIT_FILE + " " + db, os.path.join(megablastPath, tophitFound), log)
else:
log.error("- Failed to add megablast tophit file of %s." % (db))
return RQCExitCodes.JGI_FAILURE
##
## Find and add top100hit file
##
top100hitFound, _, exitCode = run_sh_command("ls %s" % (top100hitFile), True, log)
top100hitFound = top100hitFound.strip()
if top100hitFound:
append_rqc_file(filesFile, ReadqcStats.ILLUMINA_READ_TOP100HIT_FILE + " " + db, os.path.join(megablastPath, top100hitFound), log)
else:
log.error("- Failed to add megablast top100hit file of %s." % (db))
return RQCExitCodes.JGI_FAILURE
else:
log.info("- No blast hits for %s." % (db))
return RQCExitCodes.JGI_SUCCESS
"""
Title : read_level_qual_stats
Function : Generate qual scores and plots of read level 20mer sampling
Usage : read_level_mer_sampling($analysis, $summary_file_dir)
Args : 1) A reference to an JGI_Analysis object
2) current working folder wkdir/uniqueness
Returns : JGI_SUCCESS: Illumina read level report could be successfully generated.
JGI_FAILURE: Illumina read level report could not be generated.
Comments : This function is intended to be called at the very end of the illumina read level data processing script.
"""
def read_level_mer_sampling(dataToRecordDict, dataFile, log):
retCode = RQCExitCodes.JGI_FAILURE
## Old data
#nSeq nStartUniMer fracStartUniMer nRandUniMer fracRandUniMer
## 0 1 2 3 4
##25000 2500 0.1 9704 0.3882
## New data
#count first rand first_cnt rand_cnt
# 0 1 2 3 4
#25000 66.400 76.088 16600 19022
#50000 52.148 59.480 13037 14870
#75000 46.592 53.444 11648 13361
#100000 43.072 49.184 10768 12296 ...
if os.path.isfile(dataFile):
with open(dataFile, "r") as merFH:
lines = merFH.readlines()
## last line
t = lines[-1].split('\t')
# breaks 2016-09-07
#assert len(t) == 5
totalMers = int(t[0])
## new by bbcountunique
uniqStartMerPer = float("%.2f" % (float(t[1])))
uniqRandtMerPer = float("%.2f" % (float(t[2])))
dataToRecordDict[ReadqcStats.ILLUMINA_READ_20MER_SAMPLE_SIZE] = totalMers
dataToRecordDict[ReadqcStats.ILLUMINA_READ_20MER_PERCENTAGE_STARTING_MERS] = uniqStartMerPer
dataToRecordDict[ReadqcStats.ILLUMINA_READ_20MER_PERCENTAGE_RANDOM_MERS] = uniqRandtMerPer
retCode = RQCExitCodes.JGI_SUCCESS
else:
log.error("- qhist file not found: %s" % (dataFile))
return retCode
"""
Title : base_level_qual_stats
Function : Generate qual scores and plots of read level QC
Usage : base_level_qual_stats($analysis, $)
Args : 1) A reference to an JGI_Analysis object
2) current working folder wkdir/qual
Returns : JGI_SUCCESS: Illumina read level report could be successfully generated.
JGI_FAILURE: Illumina read level report could not be generated.
Comments : This function is intended to be called at the very end of the illumina base level data processing script.
"""
def base_level_qual_stats(dataToRecordDict, reformatObqhistFile, log):
cummlatPer = 0
cummlatBase = 0
statsPerc = {30:0, 25:0, 20:0, 15:0, 10:0, 5:0}
statsBase = {30:0, 25:0, 20:0, 15:0, 10:0, 5:0}
Q30_seen = 0
Q25_seen = 0
Q20_seen = 0
Q15_seen = 0
Q10_seen = 0
Q5_seen = 0
## New format
##Median 38
##Mean 37.061
##STDev 4.631
##Mean_30 37.823
##STDev_30 1.699
##Quality bases fraction
#0 159 0.00008
#1 0 0.00000
#2 12175 0.00593
#3 0 0.00000
#4 0 0.00000
#5 0 0.00000
#6 0 0.00000
allLines = open(reformatObqhistFile).readlines()
for l in allLines[::-1]:
l = l.strip()
##
## obqhist file format example
##
# #Median 36
# #Mean 33.298
# #STDev 5.890
# #Mean_30 35.303
# #STDev_30 1.517
# #Quality bases fraction
# 0 77098 0.00043
# 1 0 0.00000
# 2 0 0.00000
# 3 0 0.00000
# 4 0 0.00000
# 5 0 0.00000
# 6 0 0.00000
if len(l) > 0:
if l.startswith("#"):
if l.startswith("#Mean_30"):
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_Q30_SCORE_MEAN] = l.split('\t')[1]
elif l.startswith("#STDev_30"):
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_Q30_SCORE_STD] = l.split('\t')[1]
elif l.startswith("#Mean"):
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_OVERALL_BASES_Q_SCORE_MEAN] = l.split('\t')[1]
elif l.startswith("#STDev"):
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_OVERALL_BASES_Q_SCORE_STD] = l.split('\t')[1]
continue
qavg = None
nbase = None
percent = None
t = l.split()
try:
qavg = int(t[0])
nbase = int(t[1])
percent = float(t[2])
except IndexError:
log.warn("parse error in base_level_qual_stats: %s %s %s %s" % (l, qavg, nbase, percent))
continue
log.debug("base_level_qual_stats(): qavg and nbase and percent: %s %s %s" % (qavg, nbase, percent))
cummlatPer += percent * 100.0
cummlatPer = float("%.f" % (cummlatPer))
if cummlatPer > 100:
cummlatPer = 100.0 ## RQC-621
cummlatBase += nbase
if qavg == 30:
Q30_seen = 1
statsPerc[30] = cummlatPer
statsBase[30] = cummlatBase
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_Q30] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_C30] = cummlatBase
elif qavg == 25:
Q25_seen = 1
statsPerc[25] = cummlatPer
statsBase[25] = cummlatBase
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_Q25] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_C25] = cummlatBase
elif qavg == 20:
Q20_seen = 1
statsPerc[20] = cummlatPer
statsBase[20] = cummlatBase
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_Q20] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_C20] = cummlatBase
elif qavg == 15:
Q15_seen = 1
statsPerc[15] = cummlatPer
statsBase[15] = cummlatBase
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_Q15] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_C15] = cummlatBase
elif qavg == 10:
Q10_seen = 1
statsPerc[10] = cummlatPer
statsBase[10] = cummlatBase
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_Q10] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_C10] = cummlatBase
elif qavg == 5:
Q5_seen = 1
statsPerc[5] = cummlatPer
statsBase[5] = cummlatBase
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_Q5] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_C5] = cummlatBase
## Double check that no value is missing.
if Q25_seen == 0 and Q30_seen != 0:
Q25_seen = 1
statsPerc[25] = statsPerc[30]
statsBase[25] = statsBase[30]
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_Q25] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_C25] = cummlatBase
if Q20_seen == 0 and Q25_seen != 0:
Q20_seen = 1
statsPerc[20] = statsPerc[25]
statsBase[20] = statsBase[25]
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_Q20] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_C20] = cummlatBase
if Q15_seen == 0 and Q20_seen != 0:
Q15_seen = 1
statsPerc[15] = statsPerc[20]
statsBase[15] = statsBase[20]
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_Q15] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_C15] = cummlatBase
if Q10_seen == 0 and Q15_seen != 0:
Q10_seen = 1
statsPerc[10] = statsPerc[15]
statsBase[10] = statsBase[15]
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_Q10] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_C10] = cummlatBase
if Q5_seen == 0 and Q10_seen != 0:
Q5_seen = 1
statsPerc[5] = statsPerc[10]
statsBase[5] = statsBase[10]
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_Q5] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_BASE_C5] = cummlatBase
if Q30_seen == 0:
log.error("Q30 is 0. Base quality values are ZERO.")
log.debug("Q and C values: %s" % (dataToRecordDict))
return RQCExitCodes.JGI_SUCCESS
"""
Title : q20_score
Function : this method returns Q20 using a qrpt file as input
Usage : JGI_QC_Utility::qc20_score($qrpt)
Args : $_[0] : qrpt file.
Returns : a number of Q20 score
Comments :
"""
# def q20_score(qrpt, log):
# log.debug("qrpt file %s" % (qrpt))
#
# q20 = None
# num = 0
#
# if os.path.isfile(qrpt):
# with open(qrpt, "r") as qrptFH:
# for l in qrptFH:
# num += 1
#
# if num == 1:
# continue
#
# ##############
# ## Old format
# ## READ1.qrpt
# ## column count min max sum mean Q1 med Q3 IQR lW rW A_Count C_Count G_Count T_Count N_Count Max_count
# ## 1 378701 2 34 12447306 32.87 31 34 34 3 27 34 108573 83917 81999 104127 85 378701
# ## 2 378701 2 34 12515957 33.05 33 34 34 1 32 34 112178 83555 84449 98519 0 378701
# ## 3 378701 2 34 12519460 33.06 33 34 34 1 32 34 104668 72341 80992 120700 0 378701
# ## 4 378701 2 37 13807944 36.46 37 37 37 0 37 37 96935 95322 83958 102440 46 378701
# ## 5 378701 2 37 13790443 36.42 37 37 37 0 37 37 114586 68297 78020 117740 58 378701
# ##
# ## or
# ##
# ## READ2.qrpt
# ## column count min max sum mean Q1 med Q3 IQR lW rW A_Count C_Count G_Count T_Count N_Count Max_count
# ## 1 378701 2 34 8875097 23.44 25 26 28 3 21 32 106904 84046 81795 105956 0 378701
# ## 2 378701 2 34 6543224 17.28 15 16 26 11 2 34 107573 77148 97953 88998 7029 378701
# ## 3 378701 2 34 7131741 18.83 16 16 26 10 2 34 96452 83003 107891 91355 0 378701
# ## 4 378701 2 37 9686653 25.58 19 32 33 14 2 37 97835 78304 87944 114618 0 378701
# ## 5 378701 2 37 10208226 26.96 25 33 35 10 10 37 98021 90611 89040 101029 0 378701
#
# pos = None
# mean = None
# t = l.split("\t")
# assert len(t) > 6
# pos = int(t[0])
# mean = float(t[5])
#
# if mean and pos:
# if mean < 20:
# return pos - 1
# else:
# q20 = pos
#
# else:
# log.error("- qhist file not found: %s" % (qrpt))
# return None
#
#
# return q20
def q20_score_new(bqHist, readNum, log):
log.debug("q20_score_new(): bqHist file = %s" % (bqHist))
q20 = None
if os.path.isfile(bqHist):
with open(bqHist, "r") as qrptFH:
for l in qrptFH:
if l.startswith('#'):
continue
## New data
# 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18
##BaseNum count_1 min_1 max_1 mean_1 Q1_1 med_1 Q3_1 LW_1 RW_1 count_2 min_2 max_2 mean_2 Q1_2 med_2 Q3_2 LW_2 RW_2
# 0 6900 0 36 33.48 33 34 34 29 36 6900 0 36 33.48 33 34 34 29 36
pos = None
mean = None
t = l.split("\t")
pos = int(t[0]) + 1
if readNum == 1:
mean = float(t[4])
else:
mean = float(t[13])
if mean and pos:
if mean < 20:
return pos - 1
else:
q20 = pos
else:
log.error("- bqHist file not found: %s" % (bqHist))
return None
return q20
"""
Title : read_level_qual_stats
Function : Generate qual scores and plots of read level QC
Usage : read_level_qual_stats($analysis, $)
Args : 1) A reference to an JGI_Analysis object
2) current working folder wkdir/qual
Returns : JGI_SUCCESS: Illumina read level report could be successfully generated.
JGI_FAILURE: Illumina read level report could not be generated.
Comments : This function is intended to be called at the very end of the illumina read level data processing script.
"""
def read_level_qual_stats(dataToRecordDict, qhistTxtFullPath, log):
retCode = RQCExitCodes.JGI_FAILURE
cummlatPer = 0.0
Q30_seen = 0
Q25_seen = 0
Q20_seen = 0
Q15_seen = 0
Q10_seen = 0
Q5_seen = 0
if os.path.isfile(qhistTxtFullPath):
stats = {30:0, 25:0, 20:0, 15:0, 10:0, 5:0}
allLines = open(qhistTxtFullPath).readlines()
for l in allLines[::-1]:
if not l:
break
if l.startswith('#'):
continue
t = l.split()
assert len(t) == 3
qavg = int(t[0])
percent = float(t[2]) * 100.0 ## 20140826 Changed for bbtools
cummlatPer = cummlatPer + percent
cummlatPer = float("%.2f" % cummlatPer)
if qavg <= 30 and qavg > 25 and Q30_seen == 0:
Q30_seen = 1
stats[30] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_READ_Q30] = cummlatPer
elif qavg <= 25 and qavg > 20 and Q25_seen == 0:
Q25_seen = 1
stats[25] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_READ_Q25] = cummlatPer
elif qavg <= 20 and qavg > 15 and Q20_seen == 0:
Q20_seen = 1
stats[20] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_READ_Q20] = cummlatPer
elif qavg <= 15 and qavg > 10 and Q15_seen == 0:
Q15_seen = 1
stats[15] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_READ_Q15] = cummlatPer
elif qavg <= 10 and qavg > 5 and Q10_seen == 0:
Q10_seen = 1
stats[10] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_READ_Q10] = cummlatPer
elif qavg <= 5 and Q5_seen == 0:
Q5_seen = 1
stats[5] = cummlatPer
dataToRecordDict[ReadqcStats.ILLUMINA_READ_Q5] = cummlatPer
### Double check that no value is missing.
if Q25_seen == 0 and Q30_seen != 0:
Q25_seen = 1
stats[25] = stats[30]
dataToRecordDict[ReadqcStats.ILLUMINA_READ_Q25] = cummlatPer
if Q20_seen == 0 and Q25_seen != 0:
Q20_seen = 1
stats[20] = stats[25]
dataToRecordDict[ReadqcStats.ILLUMINA_READ_Q20] = cummlatPer
if Q15_seen == 0 and Q20_seen != 0:
Q15_seen = 1
stats[15] = stats[20]
dataToRecordDict[ReadqcStats.ILLUMINA_READ_Q15] = cummlatPer
if Q10_seen == 0 and Q15_seen != 0:
Q10_seen = 1
stats[10] = stats[15]
dataToRecordDict[ReadqcStats.ILLUMINA_READ_Q10] = cummlatPer
if Q5_seen == 0 and Q10_seen != 0:
Q5_seen = 1
stats[5] = stats[10]
dataToRecordDict[ReadqcStats.ILLUMINA_READ_Q5] = cummlatPer
if Q30_seen == 0:
log.error("Q30 is 0 . Read quality values are ZERO.")
log.debug("Q30 %s, Q25 %s, Q20 %s, Q15 %s, Q10 %s, Q5 %s" % \
(stats[30], stats[25], stats[20], stats[15], stats[10], stats[5]))
retCode = RQCExitCodes.JGI_SUCCESS
else:
log.error("- qhist file not found: %s" % (qhistTxtFullPath))
return retCode
"""
Title : read_gc_mean
Function : This function generates average GC content % and its standard deviation and put them into database.
Usage : read_gc_mean($analysis)
Args : 1) A reference to an JGI_Analysis object
Returns : JGI_SUCCESS:
JGI_FAILURE:
Comments :
"""
def read_gc_mean(histFile, log):
mean = 0.0
stdev = 0.0
retCode = RQCExitCodes.JGI_FAILURE
if os.path.isfile(histFile):
with open(histFile, "r") as histFH:
line = histFH.readline() ## we only need the first line
# Ex) #Found 1086 total values totalling 420.3971. <0.387106 +/- 0.112691>
if len(line) == 0 or not line.startswith("#Found"):
log.error("- GC content hist text file does not contains right results: %s, %s" % (histFile, line))
retCode = RQCExitCodes.JGI_FAILURE
else:
toks = line.split()
assert len(toks) == 9
mean = float(toks[6][1:]) * 100.0
stdev = float(toks[8][:-1]) * 100.0
log.debug("mean, stdev = %.2f, %.2f" % (mean, stdev))
retCode = RQCExitCodes.JGI_SUCCESS
else:
log.error("- gc hist file not found: %s" % (histFile))
return retCode, mean, stdev
if __name__ == "__main__":
exit(0)
## EOF
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