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Hi, I am trying to insert ~120 mutations to my BAM file using BAMSurgeon addSNV.py script, but I keep hitting the error:
2022-08-25T11:00:28.235353426Z [main_samview] fail to read the header from "addsnv.tmp/haplo_EB0001_1008267_1008267.tmpbam.a72f8dcd-f432-4b57-abfe-b2055ab518fe.bam.realign.sam".
2022-08-25T11:00:28.239915749Z INFO 2022-08-25 11:00:28,239 haplo_EB0001_2724975_2724975 creating tmp bam: addsnv.tmp/haplo_EB0001_2724975_2724975.tmpbam.b95d3939-19c5-464b-a6aa-6eee951a0d9a.bam
2022-08-25T11:00:28.365411158Z [Thu Aug 25 11:00:28 UTC 2022] picard.sam.SamToFastq done. Elapsed time: 0.01 minutes.
2022-08-25T11:00:28.365631285Z Runtime.totalMemory()=517996544
2022-08-25T11:00:28.408366877Z 11:00:28.406 INFO NativeLibraryLoader - Loading libgkl_compression.so from jar:file:/opt/picard.jar!/com/intel/gkl/native/libgkl_compression.so
2022-08-25T11:00:28.427371990Z INFO 2022-08-25 11:00:28,427 haplo_EB0001_220985_220985 aligning addsnv.tmp/haplo_EB0001_220985_220985.tmpbam.d7fd2714-6138-47d4-8a0e-f4527625c8ec.fastq with bwa mem
2022-08-25T11:00:28.436464771Z [E::bwa_idx_load_from_disk] fail to locate the index files
2022-08-25T11:00:28.436903841Z INFO 2022-08-25 11:00:28,436 haplo_EB0001_220985_220985 writing addsnv.tmp/haplo_EB0001_220985_220985.tmpbam.d7fd2714-6138-47d4-8a0e-f4527625c8ec.bam.realign.sam to BAM...
2022-08-25T11:00:28.441893491Z INFO 2022-08-25 11:00:28 SamToFastq
2022-08-25T11:00:28.441916700Z
2022-08-25T11:00:28.441921046Z ********** NOTE: Picard's command line syntax is changing.
2022-08-25T11:00:28.441924895Z **********
2022-08-25T11:00:28.441928440Z ********** For more information, please see:
2022-08-25T11:00:28.441932050Z **********
2022-08-25T11:00:28.441935848Z https://github.com/broadinstitute/picard/wiki/Command-Line-Syntax-Transition-For-Users-(Pre-Transition)
2022-08-25T11:00:28.441939934Z **********
2022-08-25T11:00:28.441943502Z ********** The command line looks like this in the new syntax:
2022-08-25T11:00:28.441947100Z **********
2022-08-25T11:00:28.441951172Z ********** SamToFastq -VALIDATION_STRINGENCY SILENT -INPUT addsnv.tmp/haplo_EB0001_1469895_1469895.tmpbam.843edc09-803a-4f9e-87f2-8003be7fcf34.bam -INCLUDE_NON_PRIMARY_ALIGNMENTS false -FASTQ addsnv.tmp/haplo_EB0001_1469895_1469895.tmpbam.843edc09-803a-4f9e-87f2-8003be7fcf34.fastq -INTERLEAVE true
2022-08-25T11:00:28.441956962Z **********
2022-08-25T11:00:28.441960501Z
2022-08-25T11:00:28.441963877Z
2022-08-25T11:00:28.463290448Z [main_samview] fail to read the header from "addsnv.tmp/haplo_EB0001_220985_220985.tmpbam.d7fd2714-6138-47d4-8a0e-f4527625c8ec.bam.realign.sam".
2022-08-25T11:00:28.465594101Z [Thu Aug 25 11:00:28 UTC 2022] SamToFastq INPUT=addsnv.tmp/haplo_EB0001_1666736_1666736.tmpbam.e2df6f83-9763-49c7-a1b6-71f2cf77e6b4.bam FASTQ=addsnv.tmp/haplo_EB0001_1666736_1666736.tmpbam.e2df6f83-9763-49c7-a1b6-71f2cf77e6b4.fastq INTERLEAVE=true INCLUDE_NON_PRIMARY_ALIGNMENTS=false VALIDATION_STRINGENCY=SILENT OUTPUT_PER_RG=false COMPRESS_OUTPUTS_PER_RG=false RG_TAG=PU RE_REVERSE=true INCLUDE_NON_PF_READS=false CLIPPING_MIN_LENGTH=0 READ1_TRIM=0 READ2_TRIM=0 VERBOSITY=INFO QUIET=false COMPRESSION_LEVEL=5 MAX_RECORDS_IN_RAM=500000 CREATE_INDEX=false CREATE_MD5_FILE=false GA4GH_CLIENT_SECRETS=client_secrets.json USE_JDK_DEFLATER=false USE_JDK_INFLATER=false
2022-08-25T11:00:28.467823955Z concurrent.futures.process._RemoteTraceback:
2022-08-25T11:00:28.467844140Z """
2022-08-25T11:00:28.467848901Z Traceback (most recent call last):
2022-08-25T11:00:28.467852780Z File "/usr/local/lib/python3.9/concurrent/futures/process.py", line 243, in _process_worker
2022-08-25T11:00:28.467856982Z r = call_item.fn(*call_item.args, **call_item.kwargs)
2022-08-25T11:00:28.467860852Z File "/opt/bamsurgeon/bin/addsnv.py", line 243, in makemut
2022-08-25T11:00:28.467865608Z aligners.remap_bam(args.aligner, tmpoutbamname, args.refFasta, alignopts, threads=int(args.alignerthreads), mutid=hapstr, paired=(not args.single), picardjar=args.picardjar, insane=args.insane)
2022-08-25T11:00:28.467869938Z File "/opt/bamsurgeon/bin/bamsurgeon/aligners.py", line 76, in remap_bam
2022-08-25T11:00:28.467873864Z remap_bwamem_bam(bamfn, threads, fastaref, picardjar, mutid=mutid, paired=paired, insane=insane)
2022-08-25T11:00:28.467877605Z File "/opt/bamsurgeon/bin/bamsurgeon/aligners.py", line 206, in remap_bwamem_bam
2022-08-25T11:00:28.467881602Z subprocess.check_call(bam_cmd)
2022-08-25T11:00:28.467885110Z File "/usr/local/lib/python3.9/subprocess.py", line 373, in check_call
2022-08-25T11:00:28.467889006Z raise CalledProcessError(retcode, cmd)
2022-08-25T11:00:28.467892697Z subprocess.CalledProcessError: Command '['samtools', 'view', '-bt', 'EB0001_Annotated_20211221_2.ABS27526.fasta.fai', '-o', 'addsnv.tmp/haplo_EB0001_220985_220985.tmpbam.d7fd2714-6138-47d4-8a0e-f4527625c8ec.bam', 'addsnv.tmp/haplo_EB0001_220985_220985.tmpbam.d7fd2714-6138-47d4-8a0e-f4527625c8ec.bam.realign.sam']' returned non-zero exit status 1.
2022-08-25T11:00:28.467897784Z """
2022-08-25T11:00:28.467901347Z
2022-08-25T11:00:28.467904935Z The above exception was the direct cause of the following exception:
2022-08-25T11:00:28.467908590Z
2022-08-25T11:00:28.467911627Z Traceback (most recent call last):
2022-08-25T11:00:28.467915253Z File "/opt/bamsurgeon/bin/addsnv.py", line 483, in <module>
2022-08-25T11:00:28.467981591Z [Thu Aug 25 11:00:28 UTC 2022] Executing as root@f962ac25120b on Linux 5.4.0-1071-aws amd64; OpenJDK 64-Bit Server VM 11.0.16+8-post-Ubuntu-0ubuntu120.04; Deflater: Intel; Inflater: Intel; Provider GCS is not available; Picard version: 2.27.4-SNAPSHOTINFO 2022-08-25 11:00:28,467 haplo_EB0001_2766544_2766544 creating tmp bam: addsnv.tmp/haplo_EB0001_2766544_2766544.tmpbam.c1d49a77-14c6-4e2a-8c80-78c91bb35fef.bam
2022-08-25T11:00:28.467988075Z
2022-08-25T11:00:28.468102237Z run()
2022-08-25T11:00:28.468278858Z File "/opt/bamsurgeon/bin/addsnv.py", line 480, in run
2022-08-25T11:00:28.468526462Z main(args)
2022-08-25T11:00:28.468536827Z File "/opt/bamsurgeon/bin/addsnv.py", line 390, in main
2022-08-25T11:00:28.468737497Z tmpbamlist = result.result()
2022-08-25T11:00:28.468754476Z File "/usr/local/lib/python3.9/concurrent/futures/_base.py", line 440, in result
2022-08-25T11:00:28.468983656Z return self.__get_result()
2022-08-25T11:00:28.468994882Z File "/usr/local/lib/python3.9/concurrent/futures/_base.py", line 389, in __get_result
2022-08-25T11:00:28.469193753Z raise self._exception
2022-08-25T11:00:28.469280963Z subprocess.CalledProcessError: Command '['samtools', 'view', '-bt', 'EB0001_Annotated_20211221_2.ABS27526.fasta.fai', '-o', 'addsnv.tmp/haplo_EB0001_220985_220985.tmpbam.d7fd2714-6138-47d4-8a0e-f4527625c8ec.bam', 'addsnv.tmp/haplo_EB0001_220985_220985.tmpbam.d7fd2714-6138-47d4-8a0e-f4527625c8ec.bam.realign.sam']' returned non-zero exit status 1.
I am using the latest BAMsurgeon: 1.4.1.
Dependency versions:
samtools 1.15.1
Picard 2.27.4
This is command line: python3.9 /opt/bamsurgeon/bin/addsnv.py --picardjar /opt/picard.jar --aligner mem -o Ferm619-2.simulated.bam --alignerthreads 32 -r EB0001_Annotated_20211221_2.ABS27526.fasta -v _1_EB001_simulated_mutations_with_alt.tsv -p 32 -f Ferm619-2.bam
I checked also for coverage on these positions, and there are coverage of 122X on problematic position. Can someone point me in the direction, what to debug, what to try?
The text was updated successfully, but these errors were encountered:
Hi, I am trying to insert ~120 mutations to my BAM file using BAMSurgeon addSNV.py script, but I keep hitting the error:
I am using the latest BAMsurgeon: 1.4.1.
Dependency versions:
This is command line:
python3.9 /opt/bamsurgeon/bin/addsnv.py --picardjar /opt/picard.jar --aligner mem -o Ferm619-2.simulated.bam --alignerthreads 32 -r EB0001_Annotated_20211221_2.ABS27526.fasta -v _1_EB001_simulated_mutations_with_alt.tsv -p 32 -f Ferm619-2.bam
I checked also for coverage on these positions, and there are coverage of 122X on problematic position. Can someone point me in the direction, what to debug, what to try?
The text was updated successfully, but these errors were encountered: