A USER ERROR has occurred: Bad input: We encountered a non-standard non-IUPAC base in the provided reference: '10'
If you are seeing an error, please provide(REQUIRED) :
a) GATK version used: 4.1.4.1
b) Exact command used:
docker run -v $(pwd):/2021 broadinstitute/gatk:4.1.4.1 gatk --java-options "-Xmx8G -XX:+UseParallelGC -XX:ParallelGCThreads=4" HaplotypeCaller \
-R 2021/bioinfo/lorena/bucket/reference/hg19_v0_Homo_sapiens_assembly19_v3.fasta -I 2021/scratch/bamro/ERR2880948.bam \
-L 2021/bioinfo/lorena/bucket/reference/Homo_sapiens_assembly19_v2.interval_list \
--native-pair-hmm-threads 8 \
-G StandardAnnotation -G AS_StandardAnnotation -G StandardHCAnnotation \
-O 2021/scratch/output/$(basename $sample .bam).vcf.gz -ERC GVCF
c) Entire error log:
(base) tatianaalmeida@Auto 2021 % sh scratch/runHC.sh scratch/bamro/ERR2880948.bam
01:45:30.136 WARN GATKAnnotationPluginDescriptor - Redundant enabled annotation group (StandardAnnotation) is enabled for this tool by default
01:45:30.136 WARN GATKAnnotationPluginDescriptor - Redundant enabled annotation group (StandardHCAnnotation) is enabled for this tool by default
01:45:30.164 INFO NativeLibraryLoader - Loading libgkl_compression.so from jar:file:/gatk/gatk-package-4.1.4.1-local.jar!/com/intel/gkl/native/libgkl_compression.so
Jan 15, 2021 1:45:30 AM shaded.cloud_nio.com.google.auth.oauth2.ComputeEngineCredentials runningOnComputeEngine
INFO: Failed to detect whether we are running on Google Compute Engine.
01:45:30.297 INFO HaplotypeCaller - ------------------------------------------------------------
01:45:30.298 INFO HaplotypeCaller - The Genome Analysis Toolkit (GATK) v4.1.4.1
01:45:30.298 INFO HaplotypeCaller - For support and documentation go to https://software.broadinstitute.org/gatk/
01:45:30.298 INFO HaplotypeCaller - Executing as root@ada861b931b3 on Linux v4.19.121-linuxkit amd64
01:45:30.298 INFO HaplotypeCaller - Java runtime: OpenJDK 64-Bit Server VM v1.8.0_212-8u212-b03-0ubuntu1.16.04.1-b03
01:45:30.298 INFO HaplotypeCaller - Start Date/Time: January 15, 2021 1:45:30 AM UTC
01:45:30.298 INFO HaplotypeCaller - ------------------------------------------------------------
01:45:30.298 INFO HaplotypeCaller - ------------------------------------------------------------
01:45:30.299 INFO HaplotypeCaller - HTSJDK Version: 2.21.0
01:45:30.299 INFO HaplotypeCaller - Picard Version: 2.21.2
01:45:30.299 INFO HaplotypeCaller - HTSJDK Defaults.COMPRESSION_LEVEL : 2
01:45:30.299 INFO HaplotypeCaller - HTSJDK Defaults.USE_ASYNC_IO_READ_FOR_SAMTOOLS : false
01:45:30.299 INFO HaplotypeCaller - HTSJDK Defaults.USE_ASYNC_IO_WRITE_FOR_SAMTOOLS : true
01:45:30.300 INFO HaplotypeCaller - HTSJDK Defaults.USE_ASYNC_IO_WRITE_FOR_TRIBBLE : false
01:45:30.300 INFO HaplotypeCaller - Deflater: IntelDeflater
01:45:30.300 INFO HaplotypeCaller - Inflater: IntelInflater
01:45:30.300 INFO HaplotypeCaller - GCS max retries/reopens: 20
01:45:30.300 INFO HaplotypeCaller - Requester pays: disabled
01:45:30.301 INFO HaplotypeCaller - Initializing engine
01:45:30.639 INFO FeatureManager - Using codec IntervalListCodec to read file file:///2021/scratch/reference/gatk/Homo_sapiens_assembly19_v2.interval_list
01:45:30.684 INFO IntervalArgumentCollection - Processing 2864957093 bp from intervals
01:45:30.701 INFO HaplotypeCaller - Done initializing engine
01:45:30.702 INFO HaplotypeCallerEngine - Tool is in reference confidence mode and the annotation, the following changes will be made to any specified annotations: 'StrandBiasBySample' will be enabled. 'ChromosomeCounts', 'FisherStrand', 'StrandOddsRatio' and 'QualByDepth' annotations have been disabled
01:45:30.712 INFO HaplotypeCallerEngine - Standard Emitting and Calling confidence set to 0.0 for reference-model confidence output
01:45:30.712 INFO HaplotypeCallerEngine - All sites annotated with PLs forced to true for reference-model confidence output
01:45:30.720 INFO NativeLibraryLoader - Loading libgkl_utils.so from jar:file:/gatk/gatk-package-4.1.4.1-local.jar!/com/intel/gkl/native/libgkl_utils.so
01:45:30.721 INFO NativeLibraryLoader - Loading libgkl_pairhmm_omp.so from jar:file:/gatk/gatk-package-4.1.4.1-local.jar!/com/intel/gkl/native/libgkl_pairhmm_omp.so
01:45:30.751 INFO IntelPairHmm - Flush-to-zero (FTZ) is enabled when running PairHMM
01:45:30.751 INFO IntelPairHmm - Available threads: 6
01:45:30.751 INFO IntelPairHmm - Requested threads: 8
01:45:30.751 WARN IntelPairHmm - Using 6 available threads, but 8 were requested
01:45:30.752 INFO PairHMM - Using the OpenMP multi-threaded AVX-accelerated native PairHMM implementation
01:45:30.786 INFO ProgressMeter - Starting traversal
01:45:30.786 INFO ProgressMeter - Current Locus Elapsed Minutes Regions Processed Regions/Minute
01:45:30.848 INFO VectorLoglessPairHMM - Time spent in setup for JNI call : 0.0
01:45:30.848 INFO PairHMM - Total compute time in PairHMM computeLogLikelihoods() : 0.0
01:45:30.848 INFO SmithWatermanAligner - Total compute time in java Smith-Waterman : 0.00 sec
01:45:30.848 INFO HaplotypeCaller - Shutting down engine
[January 15, 2021 1:45:30 AM UTC] org.broadinstitute.hellbender.tools.walkers.haplotypecaller.HaplotypeCaller done. Elapsed time: 0.01 minutes.
Runtime.totalMemory()=180879360
***********************************************************************
A USER ERROR has occurred: Bad input: We encountered a non-standard non-IUPAC base in the provided reference: '10'
***********************************************************************
Set the system property GATK_STACKTRACE_ON_USER_EXCEPTION (--java-options '-DGATK_STACKTRACE_ON_USER_EXCEPTION=true') to print the stack trace.
Using GATK jar /gatk/gatk-package-4.1.4.1-local.jar
Running:
java -Dsamjdk.use_async_io_read_samtools=false -Dsamjdk.use_async_io_write_samtools=true -Dsamjdk.use_async_io_write_tribble=false -Dsamjdk.compression_level=2 -Xmx8G -XX:+UseParallelGC -XX:ParallelGCThreads=4 -jar /gatk/gatk-package-4.1.4.1-local.jar HaplotypeCaller --create-output-variant-index TRUE -R /2021/scratch/reference/gatk/hg19_v0_Homo_sapiens_assembly19_v3.fasta -I /2021/scratch/bamro/ERR2880948.bam -L /2021/scratch/reference/gatk/Homo_sapiens_assembly19_v2.interval_list --native-pair-hmm-threads 8 -G StandardAnnotation -G AS_StandardAnnotation -G StandardHCAnnotation -O /2021/scratch/output/ERR2880948.g.vcf -ERC GVCF
I'm using a mac BigSur and I tried dos2unix without success.
When I apply a search for non ATCGN it only came R and M. cat -e shows a '$' at the end of the lines.
I would like some help very much,
Regards
-
Please search for this error message on google, this issue has been resolved in this forum and others multiple times. Searching for error messages in this forum and google first is the fastest way to find a resolution :)
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