CLI Reference

pycashier

_images/pycashier.svg
Usage: pycashier [OPTIONS] COMMAND [ARGS]...

  Cash in on DNA Barcode Tags

  See `pycashier COMMAND -h` for more information.

Options:
  --version   Show the version and exit.
  -h, --help  Show this message and exit.

Commands:
  extract  extract DNA barcodes from a directory of fastq files
  merge    merge overlapping paired-end reads using fastp
  receipt  combine and summarize outputs of extract
  scrna    extract expressed DNA barcodes from scRNA-seq

pycashier extract

_images/pycashier-extract.svg
Usage: pycashier extract [OPTIONS]

  extract DNA barcodes from a directory of fastq files

  Sample names should be delimited with a ".", such as `<sample>.raw.fastq`,
  anything succeeding the first period will be ignored by `pycashier`.

  If your data is paired-end with overlapping barcodes, see `pycashier merge`.

Input/Output Options:
  -i, --input DIRECTORY     source directory containing fastq files [required]
  -o, --output DIRECTORY    output directory [default: ./outs]
  -p, --pipeline DIRECTORY  pipeline directory [default: ./pipeline]
  -s, --samples TEXT        comma separated list of samples to process

Quality (Fastp) Options:
  -q, --quality INTEGER           minimum PHRED quality for filtering reads [default: 30]
  -up, --unqualified-percent INTEGER
                                  minimum percent of bases which can be below quality threshold
                                  [default: 20]
  -fa, --fastp-args TEXT          additional arguments passed to fastp [default:
                                  --dont_eval_duplication]

Trim (Cutadapt) Options:
  -ca, --cutadapt-args TEXT       additional arguments passed to cutadapt [default: --max-n=0 -n 2
                                  --trimmed-only]
  -e, --error FLOAT               error tolerance supplied to cutadapt [default: 0.1]
  -l, --length INTEGER            target length of extracted barcode [default: 20]
  -ua, --upstream-adapter TEXT    5' sequence flanking barcode
  -da, --downstream-adapter TEXT  3' sequence flanking barcode
  --unlinked-adapters             run cutadapt using unlinked adapters
  --skip-trimming                 skip cutadapt trimming entirely and use reads as-is

Cluster (Starcode) Options:
  -r, --ratio INTEGER           ratio to use for message passing clustering [default: 3]
  -d, --distance INTEGER RANGE  levenshtein distance for clustering [default: 1; 1<=x<=8]

Filter Options:
  -fc, --filter-count INTEGER  minium nominal number of reads
  -fp, --filter-percent FLOAT  minimum percentage of total reads [default: 0.005]
  --offset INTEGER             length offset from target barcode length post-clustering [default: 1]

General Options:
  -t, --threads INTEGER          number of cpu cores to use [default: 1]
  -y, --yes                      answer yes to prompts
  -v, --verbose                  show more output, set log level to debug
  -c, --config FILE              read parameter values from config file [default: pycashier.toml]
  --save-config [explicit|full]  save current params to file specified by `--config`
  --log-file FILE                path to log file [default: <pipeline-dir>/pycashier.log]
  -h, --help                     Show this message and exit.

pycashier merge

_images/pycashier-merge.svg
Usage: pycashier merge [OPTIONS]

  merge overlapping paired-end reads using fastp

  Simple wrapper over `fastp` to combine R1 and R2 from PE fastq files.

Input/Output Options:
  -i, --input DIRECTORY     source directory containing gzipped R1 and R2 fastq files [required]
  -o, --output DIRECTORY    output directory [default: ./mergedfastqs]
  -p, --pipeline DIRECTORY  pipeline directory [default: ./pipeline]
  -s, --samples TEXT        comma separated list of samples to process

Merge Options:
  -fa, --fastp-args TEXT  additional arguments passed to fastp [default: -m -c -G -Q -L]

General Options:
  -t, --threads INTEGER          number of cpu cores to use [default: 1]
  -y, --yes                      answer yes to prompts
  -v, --verbose                  show more output, set log level to debug
  -c, --config FILE              read parameter values from config file [default: pycashier.toml]
  --save-config [explicit|full]  save current params to file specified by `--config`
  --log-file FILE                path to log file [default: <pipeline-dir>/pycashier.log]
  -h, --help                     Show this message and exit.

pycashier receipt

_images/pycashier-receipt.svg
Usage: pycashier receipt [OPTIONS]

  combine and summarize outputs of extract

Input/Output Options:
  -i, --input DIRECTORY     source directory containing sam files from scRNA-seq [default: ./outs;
                            required]
  -o, --output FILE         combined tsv of all samples found in input directory [default:
                            ./combined.tsv]
  -p, --pipeline DIRECTORY  pipeline directory [default: ./pipeline]
  -s, --samples TEXT        comma separated list of samples to process

Receipt Options:
  --no-overlap  skip per lineage overlap column

General Options:
  -v, --verbose                  show more output, set log level to debug
  -c, --config FILE              read parameter values from config file [default: pycashier.toml]
  --save-config [explicit|full]  save current params to file specified by `--config`
  --log-file FILE                path to log file [default: <pipeline-dir>/pycashier.log]
  -h, --help                     Show this message and exit.

pycashier scrna

_images/pycashier-scrna.svg
Usage: pycashier scrna [OPTIONS]

  extract expressed DNA barcodes from scRNA-seq

  Designed for interoperability with 10X scRNA-seq workflow.
  After processing samples with `cellranger` resulting
  bam files should be converted to sam files using `samtools`.

  NOTE: You can speed this up by providing a sam file with only the unmapped reads.

Input/Output Options:
  -i, --input DIRECTORY     source directory containing sam files from scRNA-seq [required]
  -o, --output DIRECTORY    output directory [default: ./outs]
  -p, --pipeline DIRECTORY  pipeline directory [default: ./pipeline]
  -s, --samples TEXT        comma separated list of samples to process

Trim (Cutadapt) Options:
  -e, --error FLOAT               error tolerance supplied to cutadapt [default: 0.1]
  -l, --length INTEGER            target length of extracted barcode [default: 20]
  -ua, --upstream-adapter TEXT    5' sequence flanking barcode
  -da, --downstream-adapter TEXT  3' sequence flanking barcode
  -ca, --cutadapt-args TEXT       additional arguments passed to cutadapt [default: --max-n=0 -n 2
                                  --trimmed-only]
  -ml, --minimum-length INTEGER   minimum length of extracted barcode [default: 10]

General Options:
  -t, --threads INTEGER          number of cpu cores to use [default: 1]
  -y, --yes                      answer yes to prompts
  -v, --verbose                  show more output, set log level to debug
  -c, --config FILE              read parameter values from config file [default: pycashier.toml]
  --save-config [explicit|full]  save current params to file specified by `--config`
  --log-file FILE                path to log file [default: <pipeline-dir>/pycashier.log]
  -h, --help                     Show this message and exit.