def MakeReference(self):
if not os.path.isfile(self.strRefFile):
with open(self.strBarcodeFile) as Barcode, \
open(self.strTargetSeqFile) as Target, \
open(self.strReferenceSeqFile) as Ref, \
open(self.strRefFile, 'w') as Output:
listBarcode = Helper.RemoveNullAndBadKeyword(Barcode)
listTarget = Helper.RemoveNullAndBadKeyword(Target)
listRef = Helper.RemoveNullAndBadKeyword(Ref)
## defensive
assert len(listBarcode) == len(listTarget) == len(listRef), 'Barcode, Target and Reference must be a same row number.'
listName = []
for strBar, strTar in zip(listBarcode, listTarget):
strBar = strBar.replace('\n', '').replace('\r', '').strip().upper()
strTar = strTar.replace('\n', '').replace('\r', '').strip().upper()
Helper.CheckIntegrity(self.strBarcodeFile, strBar) ## defensive
Helper.CheckIntegrity(self.strBarcodeFile, strTar) ## defensive
listName.append(strBar + ':' + strTar + '\n')
for i, strRow in enumerate(listRef):
strRow = strRow.replace('\r', '').strip().upper()
Output.write('>' + listName[i] + strRow + '\n')
def __init__(self, strSample, strRef, options, InstInitFolder):
UserFolderAdmin.__init__(self, strSample, strRef, options,
InstInitFolder.strLogPath)
self.strSample = strSample
self._RemoveTmpBeforStart()
self.MakeSampleFolder() ## inheritance
self.strRef = strRef
self.intCore = options.multicore
self.strGapOpen = options.gap_open
self.strGapExtend = options.gap_extend
self.strTargetWindow = options.target_window
self.strIndelCheckPos = options.indel_check_pos
self.strTargetRefAlt = options.target_ref_alt
self.strBarcodeFile = os.path.join(self.strRefDir, 'Barcode.txt')
self.strReferenceSeqFile = os.path.join(self.strRefDir,
'Reference.txt')
self.strRefFile = os.path.join(self.strRefDir, 'Reference.fa')
self.strPamSeq = options.PAM_seq
self.strPamPos = options.PAM_pos
self.strGuidePos = options.Guide_pos
Helper.MakeFolderIfNot(
'./Output/{user}/{project}/{sample}/Tmp/Alignment'.format(
user=self.strUser,
project=self.strProject,
sample=self.strSample))
def RunPipeline(**kwargs):
setGroup = set()
"""
listSamples is ...
190819_Nahye_12K_D4_D0_1-Cas9D7 Cas9D7 Ctrl
190819_Nahye_12K_D4_eCas9_Rep1-Cas9D7 Cas9D7 Exp ...
"""
for strSample in listSamples:
"""
tupSampleInfo is ... tuple instance
(190819_Nahye_12K_D4_D0_1-Cas9D7, Cas9D7, Ctrl)
"""
tupSampleInfo = Helper.SplitSampleInfo(strSample)
if not tupSampleInfo: continue
"""
strSample = 190819_Nahye_12K_D4_D0_1-Cas9D7 ... sample name
, strRef = Cas9D7 ... reference name
, strExpCtrl = Ctrl/Exp/""
"""
strSample, strRef, strExpCtrl = tupSampleInfo
setGroup.add(strExpCtrl)
"""
options ... has PAM type: Cas9 Cpf1, PAM position: Forward Reverse ...
"""
InstRunner = clsIndelSearcherRunner(strSample, strRef, options, InstInitFolder)
#"""
logging.info('SplitFile')
InstRunner.SplitFile()
logging.info('MakeReference')
InstRunner.MakeReference()
"""
Indel_searcher_crispresso_hash.py
"""
logging.info('MakeIndelSearcherCmd')
listCmd = InstRunner.MakeIndelSearcherCmd()
logging.info('RunMulticore')
RunMulticore(listCmd, options.multicore) ## from CoreSystem.py
logging.info('MakeOutput')
InstRunner.MakeOutput()
logging.info('RunIndelFreqCalculator')
InstRunner.RunIndelFreqCalculator()
#"""
if setGroup == {'EXP', 'CTRL'}:
InstRunner.IndelNormalization()
elif setGroup in [set(), set([]), set(['']), set([' '])]:
pass
else:
logging.error('The group category is not appropriate. : %s' % setGroup)
logging.error('Please make sure your project file is correct.')
logging.error('The group category must be Exp or Ctrl')
raise Exception
def SubtractIndelWithD0(dictD0IndelMerge, dictExpIndel):
"""
dictD0IndelMerge: indel proportion - dictExpIndel: indel proportion
"""
strD0SubResultDir = './Output/{user}/{project}/All_results/D0SubResult'.format(
user=strUserName, project=strProjectName)
Helper.MakeFolderIfNot(strD0SubResultDir)
for strSample, dictBarcode in dictExpIndel.items():
with open(
os.path.join(
strD0SubResultDir,
'{sample}_D0SubResult.txt').format(sample=strSample),
'w') as Output:
Output.write(
'Barcode_indel_seq\tD0_total\tD0_indel_prop\tExp_total\tExp_indel_prop\tD0_sub_indel_prop\n'
)
for strBarcode, dictCountTotalAndIndel in dictBarcode.items():
intExpTotal = dictCountTotalAndIndel['Total']
for strIndelSeq, dictCount in dictCountTotalAndIndel.items():
if strIndelSeq == 'Total': continue
try:
intD0Total = dictD0IndelMerge[strBarcode]['Total']
intD0Count = dictD0IndelMerge[strBarcode][strIndelSeq][
'IndelCount']
floD0Prop = round(intD0Count / float(intD0Total), 6)
intExpCount = dictCount['IndelCount']
floExpProp = round(intExpCount / float(intExpTotal), 6)
floSubExpIndel = floExpProp - floD0Prop
if floSubExpIndel < 0:
floSubExpIndel = 0
Output.write('\t'.join(
map(str, [
strIndelSeq, intD0Total, floD0Prop,
intExpTotal, floExpProp, floSubExpIndel
])) + '\n')
except KeyError:
intExpCount = dictCount['IndelCount']
floExpProp = round(intExpCount / float(intExpTotal), 6)
Output.write('\t'.join(
map(str, [
strIndelSeq, 'None', 'None', intExpTotal,
floExpProp, floExpProp
])) + '\n')
def MakeReference(self):
with open(self.strBarcodeFile) as Barcode, \
open(self.strReferenceSeqFile) as Ref, \
open(self.strRefFile, 'w') as Output:
listBarcode = Helper.RemoveNullAndBadKeyword(Barcode)
listRef = Helper.RemoveNullAndBadKeyword(Ref)
## defensive
assert len(listBarcode) == len(
listRef), 'Barcode and Reference must be a same row number.'
dictBarcode = {}
for strBarcode in listBarcode:
strBarcode = strBarcode.replace('\n', '').replace('\r',
'').upper()
Helper.CheckIntegrity(self.strBarcodeFile,
strBarcode) ## defensive
listBarcode = strBarcode.split(':')
strBarSample = listBarcode[0]
strBarcode = listBarcode[1]
dictBarcode[strBarSample] = strBarcode
for strRef in listRef:
strRef = strRef.replace('\n', '').replace('\r', '').upper()
Helper.CheckIntegrity(self.strBarcodeFile,
strRef) ## defensive
listRef = strRef.split(':')
strRefSample = listRef[0]
strRef = listRef[1]
try:
sBarcode = dictBarcode[strRefSample]
Output.write('%s\t%s\t%s\n' %
(strRefSample, sBarcode, strRef))
except KeyError:
logging.error('no matching')
logging.error(strRefSample, strRef)
def _RemoveTmpBeforStart(self):
strFolderPath = './Output/{user}/{project}/{sample}'.format(
user=self.strUser, project=self.strProject, sample=self.strSample)
if os.path.isdir(strFolderPath):
strCmd = 'rm -r %s' % strFolderPath
Helper.PreventFromRmMistake(strCmd) ## defensive
logging.info(
'Delete the %s folder before starting if these were existed.' %
self.strSample)
sp.call(strCmd.format(user=self.strUser,
project=self.strProject,
sample=self.strSample),
shell=True)
def RunPipeline(**kwargs):
for strSample in listSamples:
if strSample[0] == '#': continue
tupSampleInfo = Helper.SplitSampleInfo(strSample)
if not tupSampleInfo: continue
strSample, strRef, strExpCtrl = tupSampleInfo
InstBaseEdit = clsBaseEditRunner(strSample, strRef, options,
InstInitFolder)
InstBaseEdit.MakeReference()
listCmd = InstBaseEdit.MakeIndelSearcherCmd()
###print(lCmd[:5])
RunMulticore(listCmd, options.multicore) ## from CoreSystem.py
InstBaseEdit.MakeMergeTarget()
InstBaseEdit.CopyToAllResultFolder()
def Main():
logging.info('Program Start')
logging.info('Make commands for a multiple processing')
lPara = []
with open('./User/{user}/{project}.txt'.format(
user=strUser, project=strProject)) as Project_list:
for strSample in Project_list:
if strSample[0] == '#': continue
tupSampleInfo = Helper.SplitSampleInfo(strSample)
if not tupSampleInfo: continue
strSample, strRef, strExpCtrl = tupSampleInfo
strSample = strSample.replace('\n', '').replace('\r', '')
sFile_path = './Output/{user}/{project}/{sample}/Tmp/Alignment'.format(
user=strUser, project=strProject, sample=strSample)
sTotal_readcnt_path = './Output/{user}/{project}/{sample}/Tmp/All'.format(
user=strUser, project=strProject, sample=strSample)
dInput_fa = Make_ref_dict(strRef)
lPara.append(
[strSample, sFile_path, sTotal_readcnt_path, dInput_fa])
logging.info('Multiple processing Start')
p = mp.Pool(iCore)
p.map_async(Count_seq_freq, lPara).get()
logging.info('Multiple processing End')
#logging.info('Count group Start')
#Count_group()
#logging.info('Count group End')
#logging.info('Trim data Start')
#Trim_data()
#logging.info('Trim data End')
logging.info('Program End')
def RunPipeline(**kwargs):
setGroup = set()
for strSample in listSamples:
tupSampleInfo = Helper.SplitSampleInfo(strSample)
if not tupSampleInfo: continue
strSample, strRef, strExpCtrl = tupSampleInfo
setGroup.add(strExpCtrl)
InstRunner = clsIndelSearcherRunner(strSample, strRef, options,
InstInitFolder)
#"""
logging.info('SplitFile')
InstRunner.SplitFile()
logging.info('MakeReference')
InstRunner.MakeReference()
logging.info('MakeIndelSearcherCmd')
listCmd = InstRunner.MakeIndelSearcherCmd()
logging.info('RunMulticore')
RunMulticore(listCmd, options.multicore) ## from CoreSystem.py
logging.info('MakeOutput')
InstRunner.MakeOutput()
logging.info('RunIndelFreqCalculator')
InstRunner.RunIndelFreqCalculator()
#"""
if setGroup == {'EXP', 'CTRL'}:
InstRunner.IndelNormalization()
elif setGroup in [set(), set([]), set(['']), set([' '])]:
pass
else:
logging.error('The group category is not appropriate. : %s' %
setGroup)
logging.error('Please make sure your project file is correct.')
logging.error('The group category must be Exp or Ctrl')
raise Exception
def CountGroup(InstParameters):
"""
Sorting_barcode Unique_RandomBarcodeNumber_In_SortingBarcode RandomBarcode Each_RandomBarcode_read_count
TATATCATAGCGTACTCATC 8 TGCGTTTG 3
TATATCATAGCGTACTCATC 8 CGCGTTTG 3
TATATCATAGCGTACTCATC 8 TAGTTTTG 1
TATATCATAGCGTACTCATC 8 ATAGTTTG 1
"""
sHeader = ''
with open(InstParameters.strSampleList) as Sample: ## tmp input
listSample = Sample.readlines()
setGroup = set([
strRow.replace('\n', '').split('\t')[2].upper()
for strRow in listSample
])
for strGroup in setGroup:
if strGroup == 'CTRL': continue
for strRow in listSample:
if strGroup == strGroupOfSample: ## matched group names -> Sum the counts
listCol = strRow.replace('\n', '').split('\t')
strSample = listCol[0]
strRef = listCol[1]
strGroupOfSample = listCol[2]
strProjectDir = './Output/{user}/{project}'.format(
user=InstParameters.strUser,
project=InstParameters.strProject)
strGroupDir = os.path.join(strProjectDir, 'Group_result')
Helper.MakeFolderIfNot(strGroupDir)
dTotal_RandomBarcode_cnt_in_SortingBarcode = OrderedDict(
) ## ('GECKO_6367_GATCTGCTC', ['GECKO_6367', 'GATCTGCTC', 2, 156, '0.0128']),
## Unique key, only one list.
with open('{project_dir}/{sample}_all_random_barcode.txt'.
format(
project_dir=strProjectDir,
sample=strSample)) as RandomBarcode_SeqFreq:
sHeader = RandomBarcode_SeqFreq.readline()
for sRow in RandomBarcode_SeqFreq:
lCol = sRow.replace('\n', '').split('\t')
sSortingBarcode = lCol[0]
#iTotal_RandomBarcode_cnt_in_SortingBarcode = int(lCol[1])
sSorting_and_Random_barcode_seq = lCol[
0] + '_' + lCol[
2] ## Unique name : Doench2014_1000_CTCTGGGGT
iRandomBarcode_count = int(lCol[3])
lCol[3] = iRandomBarcode_count
try:
_ = dTotal_RandomBarcode_cnt_in_SortingBarcode[
sSorting_and_Random_barcode_seq]
dTotal_RandomBarcode_cnt_in_SortingBarcode[
sSorting_and_Random_barcode_seq][
3] += iRandomBarcode_count
except KeyError:
dTotal_RandomBarcode_cnt_in_SortingBarcode[
sSorting_and_Random_barcode_seq] = lCol ## initial assignment
#END for
dRecal_total_kind_of_RandomBarcode = OrderedDict()
for sSort_Rand_seq in dTotal_RandomBarcode_cnt_in_SortingBarcode: ## sSorting_and_Random_barcode_seq
sSortBarcode = sSort_Rand_seq.split('_')[0]
try:
dRecal_total_kind_of_RandomBarcode[
sSortBarcode].append(
dTotal_RandomBarcode_cnt_in_SortingBarcode[
sSort_Rand_seq])
except KeyError:
dRecal_total_kind_of_RandomBarcode[
sSortBarcode] = [
dTotal_RandomBarcode_cnt_in_SortingBarcode[
sSort_Rand_seq]
]
for sKey, llValue in dRecal_total_kind_of_RandomBarcode.items(
):
## sKey: TATATCATAGCGTACTCATC, llValue : [[TATATCATAGCGTACTCATC, 8, TGCGTTTG, 3],[],[] ...
iKind_of_RandomBarcode = len(
llValue
) ################## why do I make like this ?????
for lValue in llValue:
lValue[
1] = iKind_of_RandomBarcode ## Recal using group total cnt.
llValue = sorted(llValue,
key=lambda x: x[3],
reverse=True)
dRecal_total_kind_of_RandomBarcode[sKey] = llValue
strEachGroup = './Output/Group_result/%s' % strGroup
Helper.MakeFolderIfNot(strEachGroup)
with open(os.path.join(strEachGroup, 'Summary_all_random_barcode_in_group.txt'), 'w') as Sort_Random_cnt,\
open(os.path.join(strEachGroup, 'Summary_Unique_RandomBarcodeNumber_in_group.txt'), 'w') as Uniq_random_cnt:
Sort_Random_cnt.write(sHeader)
Uniq_random_cnt.write(
'Sorting_barcode\tUnique_RandomBarcodeNumber_In_SortingBarcode\n'
)
for sSortBarcode, llCol in dRecal_total_kind_of_RandomBarcode.items(
):
Uniq_random_cnt.write(
'\t'.join(map(str, [sSortBarcode,
len(llCol)])) + '\n')
for lCol in llCol:
Sort_Random_cnt.write(
'\t'.join(map(str, lCol)) + '\n')
def Main():
print('BaseEdit program start: %s' % datetime.now())
sCmd = (
"BaseEdit frequency analyzer\n\n./Run_BaseEdit_freq.py -t 15 -w 16-48 --indel_check_pos 39-40 --target_ref_alt A,T --PAM_seq NGG --PAM_pos 43-45 --Guide_pos 23-42"
" --gap_open -10 --gap_extend 1\n\n"
"The sequence position is the one base position (start:1)\n"
"1: Barcode\n"
"2: Base target window (end pos = PAM pos +3)\n"
"3: Indel check pos\n"
"4: PAM pos\n"
"5: Guide pos (without PAM)\n\n"
"TATCTCTATCAGCACACAAGCATGCAATCACCTTGGGTCCAAAGGTCC\n"
"<------1------><----------------2--------------->\n"
" <3> <4> \n"
" <---------5--------> \n\n")
parser = OptionParser(sCmd)
parser.add_option("-t",
"--thread",
default="1",
type="int",
dest="multicore",
help="multiprocessing number")
parser.add_option('--gap_open',
default='-10',
type='float',
dest='gap_open',
help='gap open: -100~0')
parser.add_option('--gap_extend',
default='1',
type='float',
dest='gap_extend',
help='gap extend: 1~100')
parser.add_option("-w",
"--target_window",
type="str",
dest="target_window",
help="a window size for target sequence : 20-48")
parser.add_option(
"--indel_check_pos",
type="str",
dest="indel_check_pos",
help=
"indel check position to filter : 39-40; insertion 39, deletion 39 & 40"
)
parser.add_option("--target_ref_alt",
type="str",
dest="target_ref_alt",
help="Ref 'A' is changed to Alt 'T': A,T")
parser.add_option("--PAM_seq",
type="str",
dest="PAM_seq",
help="PAM sequence: NGG, NGC ...")
parser.add_option(
"--PAM_pos",
type="str",
dest="PAM_pos",
help="PAM position range in the reference seqeunce : 43-45")
parser.add_option(
"--Guide_pos",
type="str",
dest="Guide_pos",
help="Guide position range in the reference seqeunce : 23-42")
parser.add_option('--python',
dest='python',
help='The python path including the CRISPResso2')
parser.add_option('--user',
dest='user_name',
help='The user name with no space')
parser.add_option('--project',
dest='project_name',
help='The project name with no space')
options, args = parser.parse_args()
InstInitFolder = InitialFolder(options.user_name, options.project_name,
os.path.basename(__file__))
InstInitFolder.MakeDefaultFolder()
InstInitFolder.MakeInputFolder()
InstInitFolder.MakeOutputFolder()
logging.basicConfig(
format='%(process)d %(levelname)s %(asctime)s : %(message)s',
level=logging.DEBUG,
filename=InstInitFolder.strLogPath,
filemode='a')
logging.getLogger().addHandler(logging.StreamHandler(sys.stdout))
logging.info('Program start')
if options.multicore > 15:
logging.warning('Optimal treads <= 15')
logging.info(str(options))
with open(InstInitFolder.strProjectFile) as Sample_list:
listSamples = Helper.RemoveNullAndBadKeyword(Sample_list)
strInputProject = './Input/{user}/Query/{project}'.format(
user=options.user_name, project=options.project_name)
@CheckProcessedFiles
def RunPipeline(**kwargs):
for strSample in listSamples:
if strSample[0] == '#': continue
tupSampleInfo = Helper.SplitSampleInfo(strSample)
if not tupSampleInfo: continue
strSample, strRef, strExpCtrl = tupSampleInfo
InstBaseEdit = clsBaseEditRunner(strSample, strRef, options,
InstInitFolder)
InstBaseEdit.MakeReference()
listCmd = InstBaseEdit.MakeIndelSearcherCmd()
###print(lCmd[:5])
RunMulticore(listCmd, options.multicore) ## from CoreSystem.py
InstBaseEdit.MakeMergeTarget()
InstBaseEdit.CopyToAllResultFolder()
RunPipeline(InstInitFolder=InstInitFolder,
strInputProject=strInputProject,
listSamples=listSamples,
logging=logging)
print('BaseEdit program end: %s' % datetime.now())
def Main():
parser = OptionParser('Indel search program for CRISPR CAS9 & CPF1\n<All default option> python2.7 Run_indel_searcher.py --pam_type Cas9 --pam_pos Forward')
parser.add_option('-t', '--thread', default='1', type='int', dest='multicore', help='multiprocessing number, recommendation:t<16')
parser.add_option('-c', '--chunk_number', default='400000', type='int', dest='chunk_number',
help='split FASTQ, must be multiples of 4. file size < 1G recommendation:40000, size > 1G recommendation:400000')
parser.add_option('-q', '--base_quality', default='20', dest='base_quality', help='NGS read base quality')
parser.add_option('--gap_open', default='-10', type='float', dest='gap_open', help='gap open: -100~0')
parser.add_option('--gap_extend', default='1', type='float', dest='gap_extend', help='gap extend: 1~100')
parser.add_option('-i', '--insertion_window', default='4', type='int', dest='insertion_window', help='a window size for insertions')
parser.add_option('-d', '--deletion_window', default='4', type='int', dest='deletion_window', help='a window size for deletions')
parser.add_option('--pam_type', dest='pam_type', help='PAM type: Cas9 Cpf1')
parser.add_option('--pam_pos', dest='pam_pos', help='PAM position: Forward Reverse')
parser.add_option('--python', dest='python', help='The python path including the CRISPResso2')
parser.add_option('--user', dest='user_name', help='The user name with no space')
parser.add_option('--project', dest='project_name', help='The project name with no space')
parser.add_option('--pickle', dest='pickle', default='False', help='Dont remove the pickles in the tmp folder : True, False')
parser.add_option('--split', dest='split', default='False', help='Dont remove the split files in the input folder : True, False')
parser.add_option('--classfied_FASTQ', dest='class_fastq', default='True', help='Dont remove the ClassfiedFASTQ in the tmp folder : True, False')
parser.add_option('--ednafull', dest='ednafull', help='The nucleotide alignment matrix')
options, args = parser.parse_args()
InstInitFolder = InitialFolder(options.user_name, options.project_name, os.path.basename(__file__))
InstInitFolder.MakeDefaultFolder()
InstInitFolder.MakeInputFolder()
InstInitFolder.MakeOutputFolder()
logging.basicConfig(format='%(process)d %(levelname)s %(asctime)s : %(message)s',
level=logging.DEBUG,
filename=InstInitFolder.strLogPath,
filemode='a')
logging.getLogger().addHandler(logging.StreamHandler(sys.stdout))
logging.info('Program start')
if options.multicore > 15:
logging.warning('Optimal treads <= 15')
logging.info(str(options))
"""
InstInitFolder.strProjectFile is...
./User/Nahye/2019_Nahye_Cas9D7_samples.txt in INPUT3
190819_Nahye_12K_D4_D0_1-Cas9D7 Cas9D7 Ctrl
190819_Nahye_12K_D4_eCas9_Rep1-Cas9D7 Cas9D7 Exp
190819_Nahye_12K_D4_eCas9_Rep2-Cas9D7 Cas9D7 Exp
190819_Nahye_12K_D4_evo_Rep1-Cas9D7 Cas9D7 Exp
"""
with open(InstInitFolder.strProjectFile) as Sample_list:
listSamples = Helper.RemoveNullAndBadKeyword(Sample_list)
intProjectNumInTxt = len(listSamples)
strInputProject = './Input/{user}/FASTQ/{project}'.format(user=options.user_name, project=options.project_name)
@CheckProcessedFiles
def RunPipeline(**kwargs):
setGroup = set()
"""
listSamples is ...
190819_Nahye_12K_D4_D0_1-Cas9D7 Cas9D7 Ctrl
190819_Nahye_12K_D4_eCas9_Rep1-Cas9D7 Cas9D7 Exp ...
"""
for strSample in listSamples:
"""
tupSampleInfo is ... tuple instance
(190819_Nahye_12K_D4_D0_1-Cas9D7, Cas9D7, Ctrl)
"""
tupSampleInfo = Helper.SplitSampleInfo(strSample)
if not tupSampleInfo: continue
"""
strSample = 190819_Nahye_12K_D4_D0_1-Cas9D7 ... sample name
, strRef = Cas9D7 ... reference name
, strExpCtrl = Ctrl/Exp/""
"""
strSample, strRef, strExpCtrl = tupSampleInfo
setGroup.add(strExpCtrl)
"""
options ... has PAM type: Cas9 Cpf1, PAM position: Forward Reverse ...
"""
InstRunner = clsIndelSearcherRunner(strSample, strRef, options, InstInitFolder)
#"""
logging.info('SplitFile')
InstRunner.SplitFile()
logging.info('MakeReference')
InstRunner.MakeReference()
"""
Indel_searcher_crispresso_hash.py
"""
logging.info('MakeIndelSearcherCmd')
listCmd = InstRunner.MakeIndelSearcherCmd()
logging.info('RunMulticore')
RunMulticore(listCmd, options.multicore) ## from CoreSystem.py
logging.info('MakeOutput')
InstRunner.MakeOutput()
logging.info('RunIndelFreqCalculator')
InstRunner.RunIndelFreqCalculator()
#"""
if setGroup == {'EXP', 'CTRL'}:
InstRunner.IndelNormalization()
elif setGroup in [set(), set([]), set(['']), set([' '])]:
pass
else:
logging.error('The group category is not appropriate. : %s' % setGroup)
logging.error('Please make sure your project file is correct.')
logging.error('The group category must be Exp or Ctrl')
raise Exception
#"""
RunPipeline(InstInitFolder=InstInitFolder,
strInputProject=strInputProject,
intProjectNumInTxt=intProjectNumInTxt,
listSamples=listSamples,
logging=logging)
logging.info('Program end')
def __init__(self, strSample, strRef, options, InstInitFolder):
UserFolderAdmin.__init__(self, strSample, strRef, options, InstInitFolder.strLogPath)
self.MakeSampleFolder()
self.strProjectFile = InstInitFolder.strProjectFile
self.intChunkSize = options.chunk_number
self.strQualCutoff = options.base_quality
self.intInsertionWin = options.insertion_window # Insertion window 0,1,2,3,4
self.intDeletionWin = options.deletion_window # Deletion window 0,1,2,3,4
self.strPamType = options.pam_type # CRISPR type : Cpf1(2 cleavages), Cas9(1 cleavage)
self.strPamPos = options.pam_pos # Barcode target position : Forward (barcode + target), Reverse (target + barcode)
self.strPickle = options.pickle
self.strClassFASTQ = options.class_fastq
self.strSplit = options.split
self.strLogPath = InstInitFolder.strLogPath
self.strBarcodeFile = os.path.join(self.strRefDir, 'Barcode.txt')
self.strReferenceSeqFile = os.path.join(self.strRefDir, 'Reference_sequence.txt')
self.strTargetSeqFile = os.path.join(self.strRefDir, 'Target_region.txt')
self.strRefFile = os.path.join(self.strRefDir, 'Reference.fa')
## The file name required for the user is 'B'arcode.txt but it may be written as 'b'arcode.txt by mistake.
## This part is to fix the situation as mentioned above.
if not os.path.isfile(self.strBarcodeFile):
if os.path.isfile(self.strRefDir + 'barcode.txt'):
self.strBarcodeFile = self.strRefDir + 'barcode.txt'
else:
logging.error('Barcode path is not correct, please make sure the path correctly.')
if not os.path.isfile(self.strReferenceSeqFile):
if os.path.isfile(self.strRefDir + 'reference_sequence.txt'):
self.strReferenceSeqFile = self.strRefDir + 'reference_sequence.txt'
else:
logging.error('Reference path is not correct, please make sure the path correctly.')
if not os.path.isfile(self.strTargetSeqFile):
if os.path.isfile(self.strRefDir + 'target_region.txt'):
self.strTargetSeqFile = self.strRefDir + 'target_region.txt'
else:
logging.error('Target path is not correct, please make sure the path correctly.')
self.strFastqDir = './Input/{user}/FASTQ/{project}'.format(user=self.strUser,
project=self.strProject)
## './Input/JaeWoo/FASTQ/Test_samples/Sample_1'
self.strSampleDir = os.path.join(self.strFastqDir, self.strSample)
self.strFastq_name = ''
for strFile in os.listdir(self.strSampleDir):
if os.path.isfile(self.strSampleDir + '/' + strFile) and strFile.split('.')[-1] == 'fastq':
self.strFastq_name = '.'.join(strFile.split('.')[:-1])
logging.info('File name : %s' % self.strFastq_name)
## './Input/JaeWoo/FASTQ/Test_samples/Sample_1/Fastq_file.fastq'
self.strInputFile = os.path.join(self.strSampleDir, self.strFastq_name+'.fastq')
## './Input/JaeWoo/FASTQ/Test_samples/Sample_1/Fastq_file.txt'
self.strInputList = os.path.join(self.strSampleDir, self.strFastq_name+'.txt')
## './Input/JaeWoo/FASTQ/Test_samples/Sample_1/Split_files'
self.strSplitPath = os.path.join(self.strSampleDir, 'Split_files')
Helper.MakeFolderIfNot(self.strSplitPath)
self.strPair = 'False' # FASTQ pair: True, False
def Convert_Indelsearcher_output(strSampleRefGroup):
listSampleRefGroup = strSampleRefGroup.replace('\n',
'').replace('\r',
'').split('\t')
strSample = listSampleRefGroup[0]
strRef = listSampleRefGroup[1]
print('Processing: %s, %s' % (strSample, strRef))
strBaseEditRefFolder = '../Base_edit_2/Input/{user}/Reference/{project}/{ref}'.format(
user=strUser, project=strProject, ref=strRef)
strBaseEditQueryFolder = '../Base_edit_2/Input/{user}/Query/{project}/{sample}'.format(
user=strUser, project=strProject, sample=strSample)
try:
Helper.MakeFolderIfNot(strBaseEditRefFolder)
Helper.MakeFolderIfNot(strBaseEditQueryFolder)
except OSError as e:
print(e)
pass
## BaseEdit refer format : filename, barcode, reference
ReferenceFile_in_IndelSearcher = open(
'./Input/{user}/Reference/{project}/{ref}/Reference_sequence.txt'.
format(user=strUser, project=strProject, ref=strRef))
BarcodeFile_in_IndelSearcher = open(
'./Input/{user}/Reference/{project}/{ref}/Barcode.txt'.format(
user=strUser, project=strProject, ref=strRef))
BarcodeFile_for_BaseEdit = open(
'../Base_edit_2/Input/{user}/Reference/{project}/{ref}/Barcode.txt'.
format(user=strUser, project=strProject, ref=strRef), 'w')
Reference_for_BaseEdit = open(
'../Base_edit_2/Input/{user}/Reference/{project}/{ref}/Reference.txt'.
format(user=strUser, ref=strRef, project=strProject),
'w') ## conversion target to barcode:refseq
dictBarcodeSeq = {}
for strBarcodeIndelSearcher, strReferenceIndelSearcher in zip(
BarcodeFile_in_IndelSearcher, ReferenceFile_in_IndelSearcher):
strBarcodeIndelSearcher = strBarcodeIndelSearcher.replace('\n',
'').strip()
strReferenceIndelSearcher = strReferenceIndelSearcher.replace(
'\n', '').strip()
dictBarcodeSeq[strBarcodeIndelSearcher] = []
BarcodeFile_for_BaseEdit.write(
strBarcodeIndelSearcher + ':' + strBarcodeIndelSearcher + '\n'
) ## first is filename, second is barcode. BaseEdit barcode format
Reference_for_BaseEdit.write(strBarcodeIndelSearcher + ':' +
strReferenceIndelSearcher + '\n')
ReferenceFile_in_IndelSearcher.close()
BarcodeFile_in_IndelSearcher.close()
Reference_for_BaseEdit.close()
Total_result_file = open(
'./Output/{user}/{project}/{sample}/Tmp/{sample}_Classified_Indel_barcode.fastq'
.format(user=strUser, project=strProject, sample=strSample))
intCheckTotLine = 0
intOneLineMore = 0
for i, strRow in enumerate(Total_result_file): ## for query reads
if intOneLineMore == 1:
intCheckTotLine = 0
intOneLineMore = 0
if i % 4 == 0: ## Classified_Indel_barcode has all total sequence.
strBarcode = strRow.split('Barcode_')[1].split(':')[0]
intCheckTotLine = 1
elif intCheckTotLine == 1:
dictBarcodeSeq[strBarcode].append(strRow)
intOneLineMore = 1
for strBarcode, listSeq in dictBarcodeSeq.items():
with open(
'../Base_edit_2/Input/{user}/Query/{project}/{sample}/{barcode}.txt'
.format(user=strUser,
project=strProject,
sample=strSample,
barcode=strBarcode), 'w') as Output:
Output.write(''.join(listSeq))
Total_result_file.close()