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Mutect2 error "Exception in thread "main" java.lang.OutOfMemoryError: Java heap space"

Running a Mutect2 using only the required arguments gives me OutOfMemoryError. The same happens with HaplotypeCaller. I run:

./gatk Mutect2 -R /hg19.fa -I /xx.bam -O /output.vcf.gz -tumor 01

Any thoughts what might be causing it?

Answers

  • bhanuGandhambhanuGandham Cambridge MAMember, Administrator, Broadie, Moderator admin

    @KUBN

    Please post the version of gatk you are using and the entire error log.

  • KUBNKUBN Member
    edited May 2019
    Thank you @bhanuGandham . Can I also add that I tried running with --java-options "-Xmx4g" and specifying the interval of 10kb (-L chr6:31130114-31140470), is it really possible and likely that I don't have enough memory to run this analysis?

    I tried --help, --version and --CountReads commands and all work fine.
    The version is 4.0.8.1-4 and the entire log is:
    ```
    Nadas-MacBook-Pro:gatk nadakubikova$ ./gatk HaplotypeCaller -R /Users/nadakubikova/Desktop/HumanGenome/hg19.fa -I /Users/nadakubikova/Downloads/wetransfer-7475fd/C13K_S21.bam -O /Users/nadakubikova/Desktop/HumanGenome/output.g.vcf.gz --java-options "-Xmx4g" -L chr6:31130114-31140470
    Using GATK jar /Users/nadakubikova/Fish/basic/gatk/build/libs/gatk-package-4.0.8.1-4-g1dbd042-SNAPSHOT-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 -Xmx4g -jar /Users/nadakubikova/Fish/basic/gatk/build/libs/gatk-package-4.0.8.1-4-g1dbd042-SNAPSHOT-local.jar HaplotypeCaller -R /Users/nadakubikova/Desktop/HumanGenome/hg19.fa -I /Users/nadakubikova/Downloads/wetransfer-7475fd/C13K_S21.bam -O /Users/nadakubikova/Desktop/HumanGenome/output.g.vcf.gz -L chr6:31130114-31140470
    14:56:17.074 INFO NativeLibraryLoader - Loading libgkl_compression.dylib from jar:file:/Users/nadakubikova/Fish/basic/gatk/build/libs/gatk-package-4.0.8.1-4-g1dbd042-SNAPSHOT-local.jar!/com/intel/gkl/native/libgkl_compression.dylib
    14:56:17.249 INFO HaplotypeCaller - ------------------------------------------------------------
    14:56:17.250 INFO HaplotypeCaller - The Genome Analysis Toolkit (GATK) v4.0.8.1-4-g1dbd042-SNAPSHOT
    14:56:17.250 INFO HaplotypeCaller - For support and documentation go to
    14:56:17.250 INFO HaplotypeCaller - Executing as [email protected] on Mac OS X v10.14.4 x86_64
    14:56:17.250 INFO HaplotypeCaller - Java runtime: Java HotSpot(TM) 64-Bit Server VM v1.8.0_181-b13
    14:56:17.250 INFO HaplotypeCaller - Start Date/Time: 04 May 2019 14:56:17 BST
    14:56:17.250 INFO HaplotypeCaller - ------------------------------------------------------------
    14:56:17.250 INFO HaplotypeCaller - ------------------------------------------------------------
    14:56:17.251 INFO HaplotypeCaller - HTSJDK Version: 2.16.0
    14:56:17.251 INFO HaplotypeCaller - Picard Version: 2.18.7
    14:56:17.251 INFO HaplotypeCaller - HTSJDK Defaults.COMPRESSION_LEVEL : 2
    14:56:17.251 INFO HaplotypeCaller - HTSJDK Defaults.USE_ASYNC_IO_READ_FOR_SAMTOOLS : false
    14:56:17.251 INFO HaplotypeCaller - HTSJDK Defaults.USE_ASYNC_IO_WRITE_FOR_SAMTOOLS : true
    14:56:17.251 INFO HaplotypeCaller - HTSJDK Defaults.USE_ASYNC_IO_WRITE_FOR_TRIBBLE : false
    14:56:17.251 INFO HaplotypeCaller - Deflater: IntelDeflater
    14:56:17.251 INFO HaplotypeCaller - Inflater: IntelInflater
    14:56:17.251 INFO HaplotypeCaller - GCS max retries/reopens: 20
    14:56:17.251 INFO HaplotypeCaller - Using google-cloud-java fork github.com/broadinstitute/google-cloud-java/releases/tag/0.20.5-alpha-GCS-RETRY-FIX
    14:56:17.251 INFO HaplotypeCaller - Initializing engine
    15:00:21.031 INFO HaplotypeCaller - Shutting down engine
    [04 May 2019 15:00:21 BST] org.broadinstitute.hellbender.tools.walkers.haplotypecaller.HaplotypeCaller done. Elapsed time: 4.07 minutes.
    Runtime.totalMemory()=3817865216
    Exception in thread "main" java.lang.OutOfMemoryError: GC overhead limit exceeded
    at htsjdk.samtools.SAMTextHeaderCodec$ParsedHeaderLine.<init>(SAMTextHeaderCodec.java:287)
    at htsjdk.samtools.SAMTextHeaderCodec.decode(SAMTextHeaderCodec.java:95)
    at htsjdk.samtools.reference.ReferenceSequenceFileFactory.loadDictionary(ReferenceSequenceFileFactory.java:232)
    at htsjdk.samtools.reference.AbstractFastaSequenceFile.<init>(AbstractFastaSequenceFile.java:68)
    at htsjdk.samtools.reference.AbstractIndexedFastaSequenceFile.<init>(AbstractIndexedFastaSequenceFile.java:60)
    at htsjdk.samtools.reference.IndexedFastaSequenceFile.<init>(IndexedFastaSequenceFile.java:80)
    at htsjdk.samtools.reference.IndexedFastaSequenceFile.<init>(IndexedFastaSequenceFile.java:98)
    at org.broadinstitute.hellbender.utils.fasta.CachingIndexedFastaSequenceFile.<init>(CachingIndexedFastaSequenceFile.java:98)
    at org.broadinstitute.hellbender.utils.fasta.CachingIndexedFastaSequenceFile.checkAndCreate(CachingIndexedFastaSequenceFile.java:205)
    at org.broadinstitute.hellbender.utils.fasta.CachingIndexedFastaSequenceFile.checkAndCreate(CachingIndexedFastaSequenceFile.java:183)
    at org.broadinstitute.hellbender.engine.ReferenceFileSource.<init>(ReferenceFileSource.java:37)
    at org.broadinstitute.hellbender.engine.ReferenceDataSource.of(ReferenceDataSource.java:27)
    at org.broadinstitute.hellbender.engine.GATKTool.initializeReference(GATKTool.java:364)
    at org.broadinstitute.hellbender.engine.GATKTool.onStartup(GATKTool.java:634)
    at org.broadinstitute.hellbender.engine.AssemblyRegionWalker.onStartup(AssemblyRegionWalker.java:156)
    at org.broadinstitute.hellbender.cmdline.CommandLineProgram.runTool(CommandLineProgram.java:135)
    at org.broadinstitute.hellbender.cmdline.CommandLineProgram.instanceMainPostParseArgs(CommandLineProgram.java:182)
    at org.broadinstitute.hellbender.cmdline.CommandLineProgram.instanceMain(CommandLineProgram.java:201)
    at org.broadinstitute.hellbender.Main.runCommandLineProgram(Main.java:160)
    at org.broadinstitute.hellbender.Main.mainEntry(Main.java:203)
    at org.broadinstitute.hellbender.Main.main(Main.java:289)
    Nadas-MacBook-Pro:gatk nadakubikova$
    ```

    Any insights would be greatly appreciated!

    Best wishes,
    Nada
  • bhanuGandhambhanuGandham Cambridge MAMember, Administrator, Broadie, Moderator admin

    @KUBN

    How much physical memory do you have on the machine you are running this analysis? GATK tools need about 2-3GB of memory for native libraries in addition to the -Xmx value you set.

  • KUBNKUBN Member
    @bhanuGandham

    Thanks for your comment, I have got in total 8GB of physical memory.
  • KUBNKUBN Member
    @bhanuGandham

    I made sure to free up 5GB of memory. However, I have no luck running the the analysis and I still get the same error code, matter the -Xmx value I set. The bam file is not a large one. Shouldn't specifying the interval speed up the analysis?
  • bhanuGandhambhanuGandham Cambridge MAMember, Administrator, Broadie, Moderator admin
    edited May 2019

    Hi @KUBN

    I see you have posted the error log for HaplotypeCaller. Would you please post the error log for the Mutect2 command.
    Also, I would also like to know what the depth is like and whether it's particularly high where the error occurs.

    Post edited by bhanuGandham on
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