Thank you for your response. Th issue I have with the picard.jar file is, when I try to run markduplicates like this java -jar ~/picard.jar MarkDuplicates -I alignment.sort.bam -O dedup.bam -M metrics.txt -VALIDATION_STRINGENCY LENIENT -REMOVE_DUPLICATES true I get the following errors:
16:33:11.253 INFO NativeLibraryLoader - Loading libgkl_compression.so from jar:file:/home/pt689/picard.jar!/com/intel/gkl/native/libgkl_compression.so
[Mon Oct 21 16:33:26 EDT 2024] MarkDuplicates --INPUT alignment.sort.bam --OUTPUT dedup.bam --METRICS_FILE metrics.txt --REMOVE_DUPLICATES true --VALIDATION_STRINGENCY LENIENT --MAX_SEQUENCES_FOR_DISK_READ_ENDS_MAP 50000 --MAX_FILE_HANDLES_FOR_READ_ENDS_MAP 8000 --SORTING_COLLECTION_SIZE_RATIO 0.25 --TAG_DUPLICATE_SET_MEMBERS false --REMOVE_SEQUENCING_DUPLICATES false --TAGGING_POLICY DontTag --CLEAR_DT true --DUPLEX_UMI false --FLOW_MODE false --FLOW_DUP_STRATEGY FLOW_QUALITY_SUM_STRATEGY --FLOW_USE_END_IN_UNPAIRED_READS false --FLOW_USE_UNPAIRED_CLIPPED_END false --FLOW_UNPAIRED_END_UNCERTAINTY 0 --FLOW_UNPAIRED_START_UNCERTAINTY 0 --FLOW_SKIP_FIRST_N_FLOWS 0 --FLOW_Q_IS_KNOWN_END false --FLOW_EFFECTIVE_QUALITY_THRESHOLD 15 --ADD_PG_TAG_TO_READS true --ASSUME_SORTED false --DUPLICATE_SCORING_STRATEGY SUM_OF_BASE_QUALITIES --PROGRAM_RECORD_ID MarkDuplicates --PROGRAM_GROUP_NAME MarkDuplicates --READ_NAME_REGEX <optimized capture of last three ':' separated fields as numeric values> --OPTICAL_DUPLICATE_PIXEL_DISTANCE 100 --MAX_OPTICAL_DUPLICATE_SET_SIZE 300000 --VERBOSITY INFO --QUIET false --COMPRESSION_LEVEL 5 --MAX_RECORDS_IN_RAM 500000 --CREATE_INDEX false --CREATE_MD5_FILE false --help false --version false --showHidden false --USE_JDK_DEFLATER false --USE_JDK_INFLATER false
[Mon Oct 21 16:33:37 EDT 2024] Executing as pt689@argos6 on Linux 4.4.0-173-generic amd64; Java HotSpot(TM) 64-Bit Server VM 17.0.12+8-LTS-286; Deflater: Intel; Inflater: Intel; Provider GCS is available; Picard version: Version:3.3.0
INFO 2024-10-21 16:33:38 MarkDuplicates Start of doWork freeMemory: 377720336; totalMemory: 402653184; maxMemory: 32178700288
INFO 2024-10-21 16:33:38 MarkDuplicates Reading input file and constructing read end information.
INFO 2024-10-21 16:33:38 MarkDuplicates Will retain up to 116589493 data points before spilling to disk.
[Mon Oct 21 16:33:41 EDT 2024] picard.sam.markduplicates.MarkDuplicates done. Elapsed time: 0.56 minutes.
Runtime.totalMemory()=1795162112
To get help, see http://broadinstitute.github.io/picard/index.html#GettingHelp
Exception in thread "main" java.lang.NullPointerException: Cannot invoke "htsjdk.samtools.SAMReadGroupRecord.getReadGroupId()" because the return value of "htsjdk.samtools.SAMRecord.getReadGroup()" is null
at picard.sam.markduplicates.MarkDuplicates.buildSortedReadEndLists(MarkDuplicates.java:558)
at picard.sam.markduplicates.MarkDuplicates.doWork(MarkDuplicates.java:270)
at picard.cmdline.CommandLineProgram.instanceMain(CommandLineProgram.java:281)
at picard.cmdline.PicardCommandLine.instanceMain(PicardCommandLine.java:105)
at picard.cmdline.PicardCommandLine.main(PicardCommandLine.java:115)