Hi,
I am following the GATK pipeline and after running Picard CheckIlluminaDirectory and ExtractIlluminaBarcodes successfully, I encounter a problem to generate the BAM files with Illumina BasecallsToSam.
Program versions:
java version "1.8.0_77"
picardy-tools-2.2.1My command:
java -jar $HOME/bin/picard-tools-2.2.1/picard.jar IlluminaBasecallsToSam \
B=/home/user31888/exome_run_1/Data/Intensities/BaseCalls/ \
BARCODES_DIR=/home/user31888/exome_run_1/Results/ExtractIlluminaBarcodes/ \
L=1 \
RS=76T6B76T \
LIBRARY_PARAMS=/home/user31888/exome_run_1/library_param.tab \
RUN_BARCODE=exome_run_1'library_param.tab':
BARCODE OUTPUT SAMPLE_ALIAS LIBRARY_NAME
CGATGT sample_1.bam sample_1 exome_run_1
ACAGTG sample_2.bam sample_2 exome_run_1
GCCAAT sample_3.bam sample_3 exome_run_1
CAGATC sample_4.bam sample_4 exome_run_1
CTTGTA sample_5.bam sample_5 exome_run_1
GTGAAA sample_6.bam sample_6 exome_run_1Output:
picard.illumina.IlluminaBasecallsToSam BASECALLS_DIR=/home/user31888/exome_run_1/Data/Intensities/BaseCalls/ BARCODES_DIR=/home/user31888/exome_run_1/Results/ExtractIlluminaBarcodes/ LANE=1 RUN_BARCODE=exome_run_1 READ_STRUCTURE=76T6B76T LIBRARY_PARAMS=/home/user31888/exome_run_1/library_param.tab SEQUENCING_CENTER=BI PLATFORM=illumina ADAPTERS_TO_CHECK=[INDEXED, DUAL_INDEXED, NEXTERA_V2, FLUIDIGM] NUM_PROCESSORS=0 FORCE_GC=true APPLY_EAMSS_FILTER=true MAX_READS_IN_RAM_PER_TILE=1200000 MINIMUM_QUALITY=2 INCLUDE_NON_PF_READS=true IGNORE_UNEXPECTED_BARCODES=false MOLECULAR_INDEX_TAG=RX MOLECULAR_INDEX_BASE_QUALITY_TAG=QX VERBOSITY=INFO QUIET=false VALIDATION_STRINGENCY=STRICT COMPRESSION_LEVEL=5 MAX_RECORDS_IN_RAM=500000 CREATE_INDEX=false CREATE_MD5_FILE=false GA4GH_CLIENT_SECRETS=client_secrets.json
Executing as user31888@n6 on Linux 2.6.32-504.12.2.el6.664g0000.x86_64 amd64; Java HotSpot(TM) 64-Bit Server VM 1.8.0_77-b03; Picard version: 2.2.1(0256353d7b82ebcf56297abbc510da47a2ddfc0c_1459895860) IntelDeflater
picard.illumina.IlluminaBasecallsToSam done. Elapsed time: 0.00 minutes.
Runtime.totalMemory()=995098624
To get help, see http://broadinstitute.github.io/picard/index.html#GettingHelp
Exception in thread "main" picard.PicardException: LIBRARY_PARAMS(BARCODE_PARAMS) file /home/user31888/exome_run_1/library_param.tab does not have column BARCODE or BARCODE_1.
at picard.illumina.IlluminaBasecallsToSam.populateWritersFromLibraryParams(IlluminaBasecallsToSam.java:337)
at picard.illumina.IlluminaBasecallsToSam.initialize(IlluminaBasecallsToSam.java:251)
at picard.illumina.IlluminaBasecallsToSam.doWork(IlluminaBasecallsToSam.java:229)
at picard.cmdline.CommandLineProgram.instanceMain(CommandLineProgram.java:209)
at picard.cmdline.PicardCommandLine.instanceMain(PicardCommandLine.java:95)
at picard.cmdline.PicardCommandLine.main(PicardCommandLine.java:105)
The program does not see the first column BARCODE in the 'library_param.tab' file (same error message when placing the BARCODE column at the end of the file).
Is the format of the 'library_param.tab' correct?
I tried to change the end of line of the file just in case, but still the same problem.
Is it possible that the tab character could be faulty???