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atram_reference.md

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aTRAM

This takes a query sequence and a blast database built with the atram_preprocessor.py script and builds assemblies.

If you specify more than one query sequence and/or more than one blast database then aTRAM will build one assembly for each query/blast DB pair.

NOTE: You may use a text file to hold the command-line arguments like: @/path/to/args.txt. This is particularly useful when specifying multiple blast databases or multiple query sequences.

Arguments

-h, --help

Show this help message and exit.

--version

Show program's version number and exit.

-b DB [DB ...], --blast-db DB [DB ...], --sra DB [DB ...], --db DB [DB ...], --database DB [DB ...]

This needs to match the DB prefix you entered for atram_preprocessor.py. You may repeat this argument to run the --query sequence(s) against multiple blast databases.

-q QUERY [QUERY ...], --query QUERY [QUERY ...], --target QUERY [QUERY ...], --probe QUERY [QUERY ...]

The path to the fasta file with sequences of interest. You may repeat this argument. If you do then Each --query sequence file will be run against every --blast-db.

-Q QUERY_SPLIT [QUERY_SPLIT ...], --query-split QUERY_SPLIT [QUERY_SPLIT ...], --target-split QUERY_SPLIT [QUERY_SPLIT ...] The path to the fasta file with multiple sequences of interest. This will take every sequence in the fasta file and treat it as if it were its own --query argument. So every sequence in --query-split will be run against every --blast-db.

-o OUTPUT_PREFIX, --output-prefix OUTPUT_PREFIX

This is the prefix of all of the output files. So you can identify different blast output file sets. You may include a directory as part of the prefix. aTRAM will add suffixes to differentiate output files.

-a {abyss,trinity,velvet,spades,none}, --assembler {abyss,trinity,velvet,spades,none}

Which assembler to use. Choosing "none" (the default) will do a single blast run and stop before any assembly.

-i N, --iterations N

The number of pipeline iterations. The default is "5".

-p, --protein

Are the query sequences protein? aTRAM will guess if you skip this argument.

--fraction FRACTION

Use only the specified fraction of the aTRAM database. The default is 1.0.

--cpus CPUS, --processes CPUS, --max-processes CPUS

Number of CPU processors to use. This will also be used for the assemblers when possible. We will use 8 out of 12 CPUs.

--log-file LOG_FILE

Log file (full path)".

--path PATH

If the assembler or blast you want to use is not in your $PATH then use this to prepend directories to your path.

-t DIR, --temp-dir DIR

Place temporary files in this directory. All files will be deleted after aTRAM completes. The directory must exist.

--keep-temp-dir

This flag will keep the temporary files in the --temp-dir around for debugging.

-T SECONDS, --timeout SECONDS

How many seconds to wait for an assembler before stopping the run. To wait forever set this to 0. The default is "300" (5 minutes).

--no-filter

Do not filter the assembled contigs. This will: set both the --bit-score and --contig-length to 0

--bit-score SCORE

Remove contigs that have a value less than this. The default is "70.0". This is turned off by the --no- filter argument.

--contig-length CONTIG_LENGTH, --length CONTIG_LENGTH

Remove blast hits that are shorter than this length. The default is "100". This is turned off by the --no- filter argument.

--db-gencode CODE The genetic code to use during blast runs. The default is "1".

--evalue EVALUE

The default evalue is "1e-10".

--word-size WORD_SIZE Word size for wordfinder algorithm. 'Must be >= 2.

--max-target-seqs MAX

Maximum hit sequences per shard. Default is calculated based on the available memory and the number of shards.

--batch-size BATCH_SIZE

Use this option to control blast memory usage and the concatenation of queries. Setting this value too low can degrade performance.

--no-long-reads

Do not use long reads during assembly. (Abyss, Trinity, Velvet)

--kmer KMER

k-mer size. The default is 64 for Abyss and 31 for Velvet. Note: the maximum kmer length for Velvet is 31. (Abyss, Velvet)

--mpi

Use MPI for this assembler. The assembler 'must have been compiled to use MPI. (Abyss)

--bowtie2 Use bowtie2 during assembly. (Trinity)

--max-memory MEMORY

Maximum amount of memory to use in gigabytes. We will use 12 out of 24 GB of free/unused memory. (Trinity, Spades)

--exp-coverage EXP_COVERAGE, --expected-coverage EXP_COVERAGE

The expected coverage of the region. The default is "30". (Velvet)

--ins-length INS_LENGTH

The size of the fragments used in the short-read library. The default is "300". (Velvet)

--min-contig-length MIN_CONTIG_LENGTH

The minimum contig length used by the assembler itself. The default is "100". (Velvet)

--cov-cutoff COV_CUTOFF

Read coverage cutoff value. Must be a positive float value, or "auto", or "off". The default is "off". (Spades)