I am trying to use the program Trimmomatic to removed adapter sequences from an Illumina paired-end read over a computer cluster. While I can get the program to open, it will either not acknowledge the commands I enter or will return an error message. I have tried all kinds of permutations of the input commands without success. Examples of input code and error messages are below
Code:
Code: (If I add in the command PE immediately before all other commands, the program executes and can find the fasta file containing the adapter sequences, but then searches for and cannot fund a file called 'PE')
Code:
java -classpath /filepath/Trimmomatic-0.32/trimmomatic-0.32.jar org.usadellab.trimmomatic.TrimmomaticPE \
-phred33 -trimlog /Results/log.txt \
~/filepath/data_R1.fq ~/filepath/data_R2.fq \
ILLUMINACLIP:/filepath/src/Trimmomatic-0.32/adapters/TruSeq3-PE-2.fa:2:30:10:3:"true"
Results: (the o/s seems to find and execute the software, but is not feeding in the command; I get the same result if I use the java -jar <path to trimmomatic jar> option for executing Trimmomatic)-phred33 -trimlog /Results/log.txt \
~/filepath/data_R1.fq ~/filepath/data_R2.fq \
ILLUMINACLIP:/filepath/src/Trimmomatic-0.32/adapters/TruSeq3-PE-2.fa:2:30:10:3:"true"
TrimmomaticPE [-threads <threads>] [-phred33|-phred64] [-trimlog <trimLogFile>] [-basein <inputBase> | <inputFile1> <inputFile2>] [-baseout <outputBase> | <outputFile1P> <outputFile1U> <outputFile2P> <outputFile2U>] <trimmer1>...
Code: (If I add in the command PE immediately before all other commands, the program executes and can find the fasta file containing the adapter sequences, but then searches for and cannot fund a file called 'PE')
java -classpath /filepath/trimmomatic-0.32.jar org.usadellab.trimmomatic.TrimmomaticPE \
PE -phred33 -trimlog /Results/log.txt \
~/refs/lec12/data_R1.fq ~/refs/lec12/data_R2.fq \
ILLUMINACLIP:/filepath/Trimmomatic-0.32/adapters/TruSeq3-PE-2.fa:2:30:10:3:"true"
Results: (Programs rus and finds the fasta file of adapter sequences, but then fails to execute. Why is it looking for a PE file?)PE -phred33 -trimlog /Results/log.txt \
~/refs/lec12/data_R1.fq ~/refs/lec12/data_R2.fq \
ILLUMINACLIP:/filepath/Trimmomatic-0.32/adapters/TruSeq3-PE-2.fa:2:30:10:3:"true"
TrimmomaticPE: Started with arguments: PE -phred33 -trimlog /Results/log.txt /filepath/data_R1.fq /filepath/data_R2.fq ILLUMINACLIP:/filepath/Trimmomatic-0.32/adapters/TruSeq3-PE-2.fa:2:30:10:3:true
Multiple cores found: Using 12 threads
Using PrefixPair: 'TACACTCTTTCCCTACACGACGCTCTTCCGATCT' and 'GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT'
Using Long Clipping Sequence: 'AGATCGGAAGAGCACACGTCTGAACTCCAGTCAC'
Using Long Clipping Sequence: 'TACACTCTTTCCCTACACGACGCTCTTCCGATCT'
Using Long Clipping Sequence: 'GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT'
Using Long Clipping Sequence: 'AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTA'
ILLUMINACLIP: Using 1 prefix pairs, 4 forward/reverse sequences, 0 forward only sequences, 0 reverse only sequences
Exception in thread "main" java.io.FileNotFoundException: PE (No such file or directory)
at java.io.FileInputStream.open(Native Method)
at java.io.FileInputStream.<init>(FileInputStream.java:146)
at org.usadellab.trimmomatic.fastq.FastqParser.parse(FastqParser.java:127)
at org.usadellab.trimmomatic.TrimmomaticPE.process(TrimmomaticPE.java:251)
at org.usadellab.trimmomatic.TrimmomaticPE.run(TrimmomaticPE.java:498)
at org.usadellab.trimmomatic.TrimmomaticPE.main(TrimmomaticPE.java:506)
Multiple cores found: Using 12 threads
Using PrefixPair: 'TACACTCTTTCCCTACACGACGCTCTTCCGATCT' and 'GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT'
Using Long Clipping Sequence: 'AGATCGGAAGAGCACACGTCTGAACTCCAGTCAC'
Using Long Clipping Sequence: 'TACACTCTTTCCCTACACGACGCTCTTCCGATCT'
Using Long Clipping Sequence: 'GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT'
Using Long Clipping Sequence: 'AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGTA'
ILLUMINACLIP: Using 1 prefix pairs, 4 forward/reverse sequences, 0 forward only sequences, 0 reverse only sequences
Exception in thread "main" java.io.FileNotFoundException: PE (No such file or directory)
at java.io.FileInputStream.open(Native Method)
at java.io.FileInputStream.<init>(FileInputStream.java:146)
at org.usadellab.trimmomatic.fastq.FastqParser.parse(FastqParser.java:127)
at org.usadellab.trimmomatic.TrimmomaticPE.process(TrimmomaticPE.java:251)
at org.usadellab.trimmomatic.TrimmomaticPE.run(TrimmomaticPE.java:498)
at org.usadellab.trimmomatic.TrimmomaticPE.main(TrimmomaticPE.java:506)
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