23:52:16.676 WARN HDF5SVDReadCountPanelOfNormals - Exception encountered during creation of panel of normals (java.lang.IllegalArgumentException: Sample at index 0 does not have a positive sample median.
REQUIRED for all errors and issues:
a) GATK version used:4.5.0.0
b) Exact command used:
c) Entire program log:
23:50:46.694 INFO CreateReadCountPanelOfNormals - Creating the panel of normals... 23:50:46.695 INFO HDF5SVDReadCountPanelOfNormals - Creating read-count panel of normals at /cromwell_root/cnv_somatic_panel_workflow_twist_hg38_samples_for_PONs_only_4-5-0-0_2025-03-12T15-19-42.pon.hdf5... 23:50:46.696 INFO HDF5SVDReadCountPanelOfNormals - Writing version number (7.0)... 23:50:46.699 INFO HDF5SVDReadCountPanelOfNormals - Writing command line... 23:50:46.701 INFO HDF5SVDReadCountPanelOfNormals - Writing sequence dictionary... 23:50:46.720 INFO HDF5SVDReadCountPanelOfNormals - Writing original read counts (1101325 x 287)... 23:51:00.795 INFO HDF5SVDReadCountPanelOfNormals - Writing original sample filenames (287)... 23:51:00.797 INFO HDF5SVDReadCountPanelOfNormals - Writing original intervals (1101325)... 23:51:01.025 INFO HDF5SVDReadCountPanelOfNormals - Writing GC-content annotations for original intervals (1101325)... 23:51:01.038 INFO HDF5SVDReadCountPanelOfNormals - Preprocessing and standardizing read counts... 23:51:01.039 INFO SVDDenoisingUtils - Preprocessing read counts... 23:51:01.040 INFO SVDDenoisingUtils - Transforming read counts to fractional coverage... 23:51:02.918 INFO SVDDenoisingUtils - Performing GC-bias correction... 23:51:54.222 INFO SVDDenoisingUtils - Filtering intervals with median (across samples) less than or equal to the 70.00 percentile (0.00)... 23:51:54.329 INFO SVDDenoisingUtils - After filtering, 144976 out of 1101325 intervals remain... 23:51:54.329 INFO SVDDenoisingUtils - Dividing by interval medians... 23:51:59.445 INFO SVDDenoisingUtils - Filtering samples with a fraction of zero-coverage intervals greater than or equal to 80.00 percent... 23:52:05.007 INFO SVDDenoisingUtils - After filtering, 287 out of 287 samples remain... 23:52:05.007 INFO SVDDenoisingUtils - Filtering intervals with a fraction of zero-coverage samples greater than or equal to 80.00 percent... 23:52:08.103 INFO SVDDenoisingUtils - After filtering, 144976 out of 1101325 intervals remain... 23:52:13.774 INFO SVDDenoisingUtils - Filtering samples with a median (across intervals) strictly below the 2.50 percentile (1.00) or strictly above the 97.50 percentile (1.00)... 23:52:13.776 INFO SVDDenoisingUtils - After filtering, 287 out of 287 samples remain... 23:52:14.519 INFO SVDDenoisingUtils - Heap utilization statistics [MB]: 23:52:14.521 INFO SVDDenoisingUtils - Used memory: 13347 23:52:14.521 INFO SVDDenoisingUtils - Free memory: 3692 23:52:14.522 INFO SVDDenoisingUtils - Total memory: 17040 23:52:14.522 INFO SVDDenoisingUtils - Maximum memory: 29504 23:52:14.522 INFO SVDDenoisingUtils - Performing garbage collection... 23:52:14.815 INFO SVDDenoisingUtils - Heap utilization statistics [MB]: 23:52:14.815 INFO SVDDenoisingUtils - Used memory: 5346 23:52:14.815 INFO SVDDenoisingUtils - Free memory: 11693 23:52:14.815 INFO SVDDenoisingUtils - Total memory: 17040 23:52:14.815 INFO SVDDenoisingUtils - Maximum memory: 29504 23:52:15.586 INFO SVDDenoisingUtils - 101546 zero-coverage values were imputed to the median of the non-zero values in the corresponding interval... 23:52:16.282 INFO SVDDenoisingUtils - 848 values strictly below the 0.10 percentile (0.00) or strictly above the 99.90 percentile (397857249.97) were truncated to the corresponding value... 23:52:16.282 INFO SVDDenoisingUtils - Panel read counts preprocessed. 23:52:16.282 INFO SVDDenoisingUtils - Standardizing read counts... 23:52:16.282 INFO SVDDenoisingUtils - Dividing by sample medians and transforming to log2 space... 23:52:16.676 WARN HDF5SVDReadCountPanelOfNormals - Exception encountered during creation of panel of normals (java.lang.IllegalArgumentException: Sample at index 0 does not have a positive sample median.). Attempting to delete partial output in /cromwell_root/cnv_somatic_panel_workflow_twist_hg38_samples_for_PONs_only_4-5-0-0_2025-03-12T15-19-42.pon.hdf5... 23:52:16.914 INFO SparkContext - SparkContext is stopping with exitCode 0. 23:52:16.940 INFO AbstractConnector - Stopped Spark@f08f8a9{HTTP/1.1, (http/1.1)}{0.0.0.0:4040} 23:52:16.956 INFO SparkUI - Stopped Spark web UI at http://41565fb72a46:4040 23:52:16.968 INFO MapOutputTrackerMasterEndpoint - MapOutputTrackerMasterEndpoint stopped! 23:52:17.083 INFO MemoryStore - MemoryStore cleared 23:52:17.084 INFO BlockManager - BlockManager stopped 23:52:17.088 INFO BlockManagerMaster - BlockManagerMaster stopped 23:52:17.092 INFO OutputCommitCoordinator$OutputCommitCoordinatorEndpoint - OutputCommitCoordinator stopped! 23:52:17.098 INFO SparkContext - Successfully stopped SparkContext 23:52:17.099 INFO CreateReadCountPanelOfNormals - Shutting down engine [March 13, 2025 at 11:52:17 PM GMT] org.broadinstitute.hellbender.tools.copynumber.CreateReadCountPanelOfNormals done. Elapsed time: 3.64 minutes. Runtime.totalMemory()=17867735040 org.broadinstitute.hellbender.exceptions.GATKException: Could not create panel of normals. It may be necessary to use stricter parameters for filtering. For example, use a larger value of minimum-interval-median-percentile. at org.broadinstitute.hellbender.tools.copynumber.denoising.HDF5SVDReadCountPanelOfNormals.create(HDF5SVDReadCountPanelOfNormals.java:354) at org.broadinstitute.hellbender.tools.copynumber.CreateReadCountPanelOfNormals.runPipeline(CreateReadCountPanelOfNormals.java:290) at org.broadinstitute.hellbender.engine.spark.SparkCommandLineProgram.doWork(SparkCommandLineProgram.java:31) at org.broadinstitute.hellbender.cmdline.CommandLineProgram.runTool(CommandLineProgram.java:149) at org.broadinstitute.hellbender.cmdline.CommandLineProgram.instanceMainPostParseArgs(CommandLineProgram.java:198) at org.broadinstitute.hellbender.cmdline.CommandLineProgram.instanceMain(CommandLineProgram.java:217) at org.broadinstitute.hellbender.Main.runCommandLineProgram(Main.java:166) at org.broadinstitute.hellbender.Main.mainEntry(Main.java:209) at org.broadinstitute.hellbender.Main.main(Main.java:306) Caused by: java.lang.IllegalArgumentException: Sample at index 0 does not have a positive sample median. at org.broadinstitute.hellbender.utils.Utils.validateArg(Utils.java:798) at org.broadinstitute.hellbender.utils.param.ParamUtils.isPositive(ParamUtils.java:164) at org.broadinstitute.hellbender.tools.copynumber.denoising.SVDDenoisingUtils.lambda$divideBySampleMedianAndTransformToLog2$27(SVDDenoisingUtils.java:484) at java.base/java.util.stream.Streams$RangeIntSpliterator.forEachRemaining(Streams.java:104) at java.base/java.util.stream.IntPipeline$Head.forEach(IntPipeline.java:617) at org.broadinstitute.hellbender.tools.copynumber.denoising.SVDDenoisingUtils.divideBySampleMedianAndTransformToLog2(SVDDenoisingUtils.java:483) at org.broadinstitute.hellbender.tools.copynumber.denoising.SVDDenoisingUtils.preprocessAndStandardizePanel(SVDDenoisingUtils.java:86) at org.broadinstitute.hellbender.tools.copynumber.denoising.HDF5SVDReadCountPanelOfNormals.create(HDF5SVDReadCountPanelOfNormals.java:288) ... 8 more 23:52:17.104 INFO ShutdownHookManager - Shutdown hook called 23:52:17.105 INFO ShutdownHookManager - Deleting directory /cromwell_root/tmp.5BElSz/spark-b65e722d-d857-4b69-8755-a914cd0f7ee2 2025/03/13 23:52:20 Starting delocalization. 2025/03/13 23:52:21 Delocalization script execution started... 2025/03/13 23:52:21 Delocalizing output /cromwell_root/memory_retry_rc -> gs://fc-2afc6df7-8ede-4ade-a2c4-e7dbdabf7b47/submissions/78c9bace-aa45-4d37-8bfe-1546247e4565/CNVSomaticPanelWorkflow/e1a99089-fc6e-4ffb-90ab-c7c7e9c613a1/call-CreateReadCountPanelOfNormals/memory_retry_rc 2025/03/13 23:52:24 Delocalizing output /cromwell_root/rc -> gs://fc-2afc6df7-8ede-4ade-a2c4-e7dbdabf7b47/submissions/78c9bace-aa45-4d37-8bfe-1546247e4565/CNVSomaticPanelWorkflow/e1a99089-fc6e-4ffb-90ab-c7c7e9c613a1/call-CreateReadCountPanelOfNormals/rc 2025/03/13 23:52:25 Delocalizing output /cromwell_root/stdout -> gs://fc-2afc6df7-8ede-4ade-a2c4-e7dbdabf7b47/submissions/78c9bace-aa45-4d37-8bfe-1546247e4565/CNVSomaticPanelWorkflow/e1a99089-fc6e-4ffb-90ab-c7c7e9c613a1/call-CreateReadCountPanelOfNormals/stdout 2025/03/13 23:52:26 Delocalizing output /cromwell_root/stderr -> gs://fc-2afc6df7-8ede-4ade-a2c4-e7dbdabf7b47/submissions/78c9bace-aa45-4d37-8bfe-1546247e4565/CNVSomaticPanelWorkflow/e1a99089-fc6e-4ffb-90ab-c7c7e9c613a1/call-CreateReadCountPanelOfNormals/stderr 2025/03/13 23:52:28 Delocalizing output /cromwell_root/cnv_somatic_panel_workflow_twist_hg38_samples_for_PONs_only_4-5-0-0_2025-03-12T15-19-42.pon.hdf5 -> gs://fc-2afc6df7-8ede-4ade-a2c4-e7dbdabf7b47/submissions/78c9bace-aa45-4d37-8bfe-1546247e4565/CNVSomaticPanelWorkflow/e1a99089-fc6e-4ffb-90ab-c7c7e9c613a1/call-CreateReadCountPanelOfNormals/cnv_somatic_panel_workflow_twist_hg38_samples_for_PONs_only_4-5-0-0_2025-03-12T15-19-42.pon.hdf5 Required file output '/cromwell_root/cnv_somatic_panel_workflow_twist_hg38_samples_for_PONs_only_4-5-0-0_2025-03-12T15-19-42.pon.hdf5' does not exist.
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Hi Zunqiu Wang
Have you tried running by adjusting the minimum interval median percentile parameter to a higher value? Looks like filtering of intervals ended up zero valid results therefore it cannot perform a proper PON creation. You may want to QC your samples for PON creation before getting to this stage and see if there are bad samples that result in such error. If samples are hybrid capture in nature you can check CollectHsMetrics results to see possible outliers.
Regards.
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