# HG changeset patch # User greg # Date 1587478793 14400 # Node ID c21d338dbdc4966553ce71e5ec51042840cb2eef Uploaded diff -r 000000000000 -r c21d338dbdc4 .shed.yml --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/.shed.yml Tue Apr 21 10:19:53 2020 -0400 @@ -0,0 +1,11 @@ +name: vsnp_statistics +owner: greg +description: | + Contains a tool that outputs statistics on VCF files. +homepage_url: https://github.com/USDA-VS/vSNP +long_description: | + Contains a tool that outputs statistics on VCF files. +remote_repository_url: https://github.com/gregvonkuster/galaxy_tools/tree/master/tools/sequence_analysis/vsnp/vsnp_statistics +type: unrestricted +categories: + - Sequence Analysis diff -r 000000000000 -r c21d338dbdc4 test-data/13-1941-6_S4_L001_R1_600000.fastq.gz Binary file test-data/13-1941-6_S4_L001_R1_600000.fastq.gz has changed diff -r 000000000000 -r c21d338dbdc4 test-data/13-1941-6_S4_L001_R2_600000.fastq.gz Binary file test-data/13-1941-6_S4_L001_R2_600000.fastq.gz has changed diff -r 000000000000 -r c21d338dbdc4 test-data/add_zc_metrics.tabular --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/test-data/add_zc_metrics.tabular Tue Apr 21 10:19:53 2020 -0400 @@ -0,0 +1,3 @@ +# File Number of Good SNPs Average Coverage Genome Coverage +MarkDuplicates on data 4_ MarkDuplicates BAM output 10.338671 98.74% +VCFfilter_ on data 7 611 diff -r 000000000000 -r c21d338dbdc4 test-data/samtools_idxstats.tabular --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/test-data/samtools_idxstats.tabular Tue Apr 21 10:19:53 2020 -0400 @@ -0,0 +1,2 @@ +NC_002945.4 4349904 45 4047 +* 0 0 5 diff -r 000000000000 -r c21d338dbdc4 test-data/vsnp_statistics.tabular --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/test-data/vsnp_statistics.tabular Tue Apr 21 10:19:53 2020 -0400 @@ -0,0 +1,22 @@ +Sample 13-1941-6 +Reference 89 +Read1 File 13-1941-6_S4_L001_R1_600000_fastq_gz +Read1 File Size 5.1 KB +Read1 Total Reads 25 +Read1 Mean Read Length 230.6 +Read1 Mean Read Quality 30.8 +Read1 Reads Passing Q30 60.0% +Read2 File 13-1941-6_S4_L001_R2_600000_fastq_gz +Read2 File Size 5.6 KB +Read2 Total Reads 25 +Read2 Mean Read Length 239.7 +Read2 Mean Read Quality 22.2 +Read2 Reads Passing Q30 4.0% +Total Reads 50 +All Mapped Reads 45 +Unmapped Reads 5 +Unmapped Reads Percentage of Total 10.0% +Average Depth of Coverage 10.338671 +Reference with Coverage 98.74% + +Good SNP Count 611 diff -r 000000000000 -r c21d338dbdc4 vsnp_statistics.py --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/vsnp_statistics.py Tue Apr 21 10:19:53 2020 -0400 @@ -0,0 +1,236 @@ +#!/usr/bin/env python + +import argparse +import gzip +import multiprocessing +import numpy +import os +import pandas +import queue + +INPUT_IDXSTATS_DIR = 'input_idxstats' +INPUT_METRICS_DIR = 'input_metrics' +INPUT_READS_DIR = 'input_reads' +OUTPUT_DIR = 'output' +QUALITYKEY = {'!':'0', '"':'1', '#':'2', '$':'3', '%':'4', '&':'5', "'":'6', '(':'7', ')':'8', '*':'9', '+':'10', ',':'11', '-':'12', '.':'13', '/':'14', '0':'15', '1':'16', '2':'17', '3':'18', '4':'19', '5':'20', '6':'21', '7':'22', '8':'23', '9':'24', ':':'25', ';':'26', '<':'27', '=':'28', '>':'29', '?':'30', '@':'31', 'A':'32', 'B':'33', 'C':'34', 'D':'35', 'E':'36', 'F':'37', 'G':'38', 'H':'39', 'I':'40', 'J':'41', 'K':'42', 'L':'43', 'M':'44', 'N':'45', 'O':'46', 'P':'47', 'Q':'48', 'R':'49', 'S':'50', 'T':'51', 'U':'52', 'V':'53', 'W':'54', 'X':'55', 'Y':'56', 'Z':'57', '_':'1', ']':'1', '[':'1', '\\':'1', '\n':'1', '`':'1', 'a':'1', 'b':'1', 'c':'1', 'd':'1', 'e':'1', 'f':'1', 'g':'1', 'h':'1', 'i':'1', 'j':'1', 'k':'1', 'l':'1', 'm':'1', 'n':'1', 'o':'1', 'p':'1', 'q':'1', 'r':'1', 's':'1', 't':'1', 'u':'1', 'v':'1', 'w':'1', 'x':'1', 'y':'1', 'z':'1', ' ':'1'} +READCOLUMNS = ['Sample', 'Reference', 'Fastq File', 'Size', 'Total Reads', 'Mean Read Length', 'Mean Read Quality', 'Reads Passing Q30'] +SEP = "\t" + + +def get_base_file_name(file_path): + base_file_name = os.path.basename(file_path) + if base_file_name.find(".") > 0: + # Eliminate the extension. + return os.path.splitext(base_file_name)[0] + elif base_file_name.find("_") > 0: + # The dot extension was likely changed to + # the " character. + items = base_file_name.split("_") + return "_".join(items[0:-1]) + else: + return base_file_name + + +def nice_size(size): + # Returns a readably formatted string with the size + words = ['bytes', 'KB', 'MB', 'GB', 'TB', 'PB', 'EB'] + prefix = '' + try: + size = float(size) + if size < 0: + size = abs(size) + prefix = '-' + except Exception: + return '??? bytes' + for ind, word in enumerate(words): + step = 1024 ** (ind + 1) + if step > size: + size = size / float(1024 ** ind) + if word == 'bytes': # No decimals for bytes + return "%s%d bytes" % (prefix, size) + return "%s%.1f %s" % (prefix, size, word) + return '??? bytes' + + +def output_read_stats(gzipped, fastq_file, ofh, sampling_number=10000, output_sample=False, dbkey=None, collection=False): + file_name_base = os.path.basename(fastq_file) + # Output a 2-column file where column 1 is + # the labels and column 2 is the values. + if output_sample: + # The Sample and Reference columns should be + # output only once, so this block handles + # paired reads, where the columns are not + # output for Read2. + try: + # Illumina read file names are something like: + # 13-1941-6_S4_L001_R1_600000_fastq_gz + sample = file_name_base.split("_")[0] + except Exception: + sample = "" + # Sample + ofh.write("Sample%s%s\n" % (SEP, sample)) + ofh.write("Reference%s%s\n" % (SEP, dbkey)) + if collection: + read = "Read" + else: + read = "Read1" + else: + read = "Read2" + # Read + ofh.write("%s File%s%s\n" % (read, SEP, file_name_base)) + # File Size + ofh.write("%s File Size%s%s\n" % (read, SEP, nice_size(os.path.getsize(fastq_file)))) + if gzipped.lower() == "true": + df = pandas.read_csv(gzip.open(fastq_file, "r"), header=None, sep="^") + else: + df = pandas.read_csv(open(fastq_file, "r"), header=None, sep="^") + total_read_count = int(len(df.index) / 4) + # Readx Total Reads + ofh.write("%s Total Reads%s%s\n" % (read, SEP, total_read_count)) + # Mean Read Length + sampling_size = int(sampling_number) + if sampling_size > total_read_count: + sampling_size = total_read_count + df = df.iloc[3::4].sample(sampling_size) + dict_mean = {} + list_length = [] + for index, row in df.iterrows(): + base_qualities = [] + for base in list(row.array[0]): + base_qualities.append(int(QUALITYKEY[base])) + dict_mean[index] = numpy.mean(base_qualities) + list_length.append(len(row.array[0])) + ofh.write("%s Mean Read Length%s%s\n" % (read, SEP, "%.1f" % numpy.mean(list_length))) + # Mean Read Quality + df_mean = pandas.DataFrame.from_dict(dict_mean, orient='index', columns=['ave']) + ofh.write("%s Mean Read Quality%s%s\n" % (read, SEP, "%.1f" % df_mean['ave'].mean())) + # Reads Passing Q30 + reads_gt_q30 = len(df_mean[df_mean['ave'] >= 30]) + reads_passing_q30 = "{:10.2f}".format(reads_gt_q30 / sampling_size) + ofh.write("%s reads passing Q30%s%s\n" % (read, SEP, reads_passing_q30)) + return total_read_count + + +def output_statistics(task_queue, read2, collection, gzipped, dbkey, timeout): + while True: + try: + tup = task_queue.get(block=True, timeout=timeout) + except queue.Empty: + break + read_file, idxstats_file, metrics_file, output_file = tup + total_reads = 0 + with open(output_file, "w") as ofh: + total_reads += output_read_stats(gzipped, read_file, ofh, output_sample=True, dbkey=dbkey, collection=collection) + if read2 is not None: + total_reads += output_read_stats(gzipped, read2, ofh) + ofh.write("Total Reads%s%d\n" % (SEP, total_reads)) + with open(idxstats_file, "r") as ifh: + unmapped_reads = 0 + for i, line in enumerate(ifh): + items = line.split("\t") + if i == 0: + # NC_002945.4 4349904 213570 4047 + ofh.write("All Mapped Reads%s%s\n" % (SEP, items[2])) + elif i == 1: + # * 0 0 82774 + unmapped_reads = int(items[3]) + ofh.write("Unmapped Reads%s%d\n" % (SEP, unmapped_reads)) + percent_str = "Unmapped Reads Percentage of Total" + if unmapped_reads > 0: + unmapped_reads_percentage = "{:10.2f}".format(unmapped_reads / total_reads) + ofh.write("%s%s%s\n" % (percent_str, SEP, unmapped_reads_percentage)) + else: + ofh.write("%s%s0\n" % (percent_str, SEP)) + with open(metrics_file, "r") as ifh: + for i, line in enumerate(ifh): + if i == 0: + # Skip comments. + continue + items = line.split("\t") + if i == 1: + # MarkDuplicates 10.338671 98.74% + ofh.write("Average Depth of Coverage%s%s\n" % (SEP, items[2])) + ofh.write("Reference with Coverage%s%s\n" % (SEP, items[3])) + elif i == 2: + # VCFfilter 611 + ofh.write("Good SNP Count%s%s\n" % (SEP, items[1])) + task_queue.task_done() + + +def set_num_cpus(num_files, processes): + num_cpus = int(multiprocessing.cpu_count()) + if num_files < num_cpus and num_files < processes: + return num_files + if num_cpus < processes: + half_cpus = int(num_cpus / 2) + if num_files < half_cpus: + return num_files + return half_cpus + return processes + + +if __name__ == '__main__': + parser = argparse.ArgumentParser() + + parser.add_argument('--read1', action='store', dest='read1', required=False, default=None, help='Required: single read') + parser.add_argument('--read2', action='store', dest='read2', required=False, default=None, help='Optional: paired read') + parser.add_argument('--dbkey', action='store', dest='dbkey', help='Reference dbkey') + parser.add_argument('--gzipped', action='store', dest='gzipped', help='Input files are gzipped') + parser.add_argument('--samtools_idxstats', action='store', dest='samtools_idxstats', required=False, default=None, help='Output of samtools_idxstats') + parser.add_argument('--output', action='store', dest='output', required=False, default=None, help='Output statisticsfile') + parser.add_argument('--vsnp_azc', action='store', dest='vsnp_azc', required=False, default=None, help='Output of vsnp_add_zero_coverage') + parser.add_argument('--processes', action='store', dest='processes', type=int, help='User-selected number of processes to use for job splitting') + + args = parser.parse_args() + + reads_files = [] + idxstats_files = [] + metrics_files = [] + output_files = [] + if args.output is not None: + # The inputs were not dataset collections, so + # read1, read2 (possibly) and vsnp_azc will also + # not be None. + collection = False + reads_files.append(args.read1) + idxstats_files.append(args.samtools_idxstats) + metrics_files.append(args.vsnp_azc) + output_files.append(args.output) + else: + collection = True + for file_name in sorted(os.listdir(INPUT_READS_DIR)): + file_path = os.path.abspath(os.path.join(INPUT_READS_DIR, file_name)) + reads_files.append(file_path) + base_file_name = get_base_file_name(file_path) + output_files.append(os.path.abspath(os.path.join(OUTPUT_DIR, "%s.tabular" % base_file_name))) + for file_name in sorted(os.listdir(INPUT_IDXSTATS_DIR)): + file_path = os.path.abspath(os.path.join(INPUT_IDXSTATS_DIR, file_name)) + idxstats_files.append(file_path) + for file_name in sorted(os.listdir(INPUT_METRICS_DIR)): + file_path = os.path.abspath(os.path.join(INPUT_METRICS_DIR, file_name)) + metrics_files.append(file_path) + + multiprocessing.set_start_method('spawn') + queue1 = multiprocessing.JoinableQueue() + num_files = len(output_files) + cpus = set_num_cpus(num_files, args.processes) + # Set a timeout for get()s in the queue. + timeout = 0.05 + + for i, output_file in enumerate(output_files): + read_file = reads_files[i] + idxstats_file = idxstats_files[i] + metrics_file = metrics_files[i] + queue1.put((read_file, idxstats_file, metrics_file, output_file)) + + # Complete the output_statistics task. + processes = [multiprocessing.Process(target=output_statistics, args=(queue1, args.read2, collection, args.gzipped, args.dbkey, timeout, )) for _ in range(cpus)] + for p in processes: + p.start() + for p in processes: + p.join() + queue1.join() + + if queue1.empty(): + queue1.close() + queue1.join_thread() diff -r 000000000000 -r c21d338dbdc4 vsnp_statistics.xml --- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/vsnp_statistics.xml Tue Apr 21 10:19:53 2020 -0400 @@ -0,0 +1,154 @@ + + + + humanize + numpy + pandas + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + input_type_cond['input_type'] == 'single' + + + + input_type_cond['input_type'] == 'collection' + + + + + + + + + + + + +**What it does** + +Accepts a single fastqsanger read, a set of paired reads, or a collections of reads along with associated SAMtools +idxstats and vSNP zero coverage metrics files and extracts information from the files to produce a tabular statistics +dataset that includes total reads, mean read length and quality, reads passing Q30, mapped and unmapped reads, depth +of coverage, good SNP count and more. + +**Required options** + + * **Choose the type for files to be analyzed** - select "Single files" or "Collections of files", then select the appropriate history items (single or paired fastqsanger reads or collections of fastqsanger reads and associated idxstats and vSNP zero coverage metrics files) based on the selected option.. + * **Number of processes for job splitting** - Select the number of processes for splitting the job to shorten execution time. + + + + @misc{None, + journal = {None}, + author = {1. Stuber T}, + title = {Manuscript in preparation}, + year = {None}, + url = {https://github.com/USDA-VS/vSNP},} + + + +