1st Attempt:
cat ./*_R1_001.fastq > 5RM1_R1.fastq
...
cd /project/microbiome/data_queue/seq/loc_ad1/otu
./run_slurm_mkotu.pl
Just analyzing trimmed R1s to avoid suspected merge/join bias because of 2 x 150 sequencing:
cd /project/microbiome/data_queue/seq/TRNLloc_Testad1/tfmergedreads/16S/TRNL1loc_ad1/trimmed
cp *./*R1.fq /project/microbiome/data_queue/seq/TRNLloc_Testad1/R1only/
cd /project/microbiome/data_queue/seq/TRNLloc_Testad1/R1only
sed -n '1~4s/^@/>/p;2~4p' ./*.fq > ./TRNL_Test_16SLocAdR1.fa
vsearch -derep_fulllength --input TrnlTest16S.fa --output uniqueSequences.fa --sizeout --sizein
Code Block |
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vsearch --derep_fulllength $s |
...
LocAdR1.fa \ --strand plus \ --output $s derep.fa \ --sizeout \ --uc $s.derep.uc \ --relabel $s. \ --fasta_width 0 |
vsearch --cluster_unoise derep.fa --centroids zotus_vsearch.fa --sizein --sizeout
3rd Attempt:
Reran 1st attempt commands starting with “./run_slurm_mergereads.pl
“
Edited “215” in the below code chunk of trim_merge.pl to “115”
Code Block |
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system("vsearch --fastx_filter $R1tmpA --fastq_trunclen 215 --fastqout $R1tmpB --threads 32"); system("vsearch --fastx_filter $R2tmpA --fastq_trunclen 215 --fastqout $R2tmpB --threads 32"); print "vsearch step1 complete\n"; if(-e $R1tmpB && -e $R1tmpB){ system("vsearch --fastq_join $R1tmpB --reverse $R2tmpB --fastaout $joinedfile --threads 32"); unlink($R1tmpA, $R2tmpA, $R1tmpB, $R2tmpB); } |