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Stopped here on 9-01-22

 

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Gregg Randolph : please see /project/gtl/data/raw/ALF1/16S/tfmergedreads where I made mergereads.nf, teton.conf and edited trim_merge.pl to trim_mergecab.pl (initially this was because I didn't have permissions to run the file, so I copied it, but I found I need to make some changes, with are in the *cab.pl version).  You run the nextflow script with: module load nextflowand nextflow run -bg mergereads.nf  -c teton.config. See inside of mergereads.nf for other ways of running it (i.e., not in the background). I tried this on pair of input files and one of the vsearch steps in the middle fails because the inputs are too small. But the nextflow script completes. Please have a look and see what you find and can figure out. It might be that some of the input files are genuinely too small.

I can see that the trimming step is working. You can see this in output/trimmed/. But we're not getting other files (in unmerged/, joined/, and in output/ itself.).

Logs and other files for debugging will be typically be in work/ For example, see work/ff/d9576360a5295e6c92a0183e485944/.command.log and neighboring files with ls -al work/ff/d9576360a5295e6c92a0183e485944/. In this case I think vsearch is silently failing to write anything. Importantly, you can see the command that was being run, to debug: see work/ff/d9576360a5295e6c92a0183e485944/.command.sh

Trim, merge and filter reads

In /project/gtl/data/raw/ALF1/16S/rawdata/tfmergedreads , we used run_slurm_mergereads.plto crawl the project folders and sample files (created in the splitting step above) to merge read pairs, and filter based on base quality. This script conforms to the steps in https://microcollaborative.atlassian.net/wiki/spaces/MICLAB/pages/1123778569/Bioinformatics+v3.0?focusedCommentId=1280377080#comment-1280377080, including trimming primers, and joining unmerged reads. This writes a new set of fasta files for each sample and project, rather than fastq, to be used in subsequent steps. These files are found in the 16S/ and ITS/ folders in tfmergedreads/. For example, see contents of /project/gtl/data/distribution/ALF1/ITS/rawdata/

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