Program options for hisat2: HISAT2 version 2.1.0 by Daehwan Kim (infphilo@gmail.com, www.ccb.jhu.edu/people/infphilo) Usage: hisat2 [options]* -x {-1 -2 | -U | --sra-acc } [-S ] Index filename prefix (minus trailing .X.ht2). Files with #1 mates, paired with files in . Could be gzip'ed (extension: .gz) or bzip2'ed (extension: .bz2). Files with #2 mates, paired with files in . Could be gzip'ed (extension: .gz) or bzip2'ed (extension: .bz2). Files with unpaired reads. Could be gzip'ed (extension: .gz) or bzip2'ed (extension: .bz2). Comma-separated list of SRA accession numbers, e.g. --sra-acc SRR353653,SRR353654. File for SAM output (default: stdout) , , can be comma-separated lists (no whitespace) and can be specified many times. E.g. '-U file1.fq,file2.fq -U file3.fq'. Options (defaults in parentheses): Input: -q query input files are FASTQ .fq/.fastq (default) --qseq query input files are in Illumina's qseq format -f query input files are (multi-)FASTA .fa/.mfa -r query input files are raw one-sequence-per-line -c , , are sequences themselves, not files -s/--skip skip the first reads/pairs in the input (none) -u/--upto stop after first reads/pairs (no limit) -5/--trim5 trim bases from 5'/left end of reads (0) -3/--trim3 trim bases from 3'/right end of reads (0) --phred33 qualities are Phred+33 (default) --phred64 qualities are Phred+64 --int-quals qualities encoded as space-delimited integers --sra-acc SRA accession ID Alignment: --n-ceil func for max # non-A/C/G/Ts permitted in aln (L,0,0.15) --ignore-quals treat all quality values as 30 on Phred scale (off) --nofw do not align forward (original) version of read (off) --norc do not align reverse-complement version of read (off) Spliced Alignment: --pen-cansplice penalty for a canonical splice site (0) --pen-noncansplice penalty for a non-canonical splice site (12) --pen-canintronlen penalty for long introns (G,-8,1) with canonical splice sites --pen-noncanintronlen penalty for long introns (G,-8,1) with noncanonical splice sites --min-intronlen minimum intron length (20) --max-intronlen maximum intron length (500000) --known-splicesite-infile provide a list of known splice sites --novel-splicesite-outfile report a list of splice sites --novel-splicesite-infile provide a list of novel splice sites --no-temp-splicesite disable the use of splice sites found --no-spliced-alignment disable spliced alignment --rna-strandness specify strand-specific information (unstranded) --tmo reports only those alignments within known transcriptome --dta reports alignments tailored for transcript assemblers --dta-cufflinks reports alignments tailored specifically for cufflinks --avoid-pseudogene tries to avoid aligning reads to pseudogenes (experimental option) --no-templatelen-adjustment disables template length adjustment for RNA-seq reads Scoring: --mp , max and min penalties for mismatch; lower qual = lower penalty <6,2> --sp , max and min penalties for soft-clipping; lower qual = lower penalty <2,1> --no-softclip no soft-clipping --np penalty for non-A/C/G/Ts in read/ref (1) --rdg , read gap open, extend penalties (5,3) --rfg , reference gap open, extend penalties (5,3) --score-min min acceptable alignment score w/r/t read length (L,0.0,-0.2) Reporting: -k (default: 5) report up to alns per read Paired-end: -I/--minins minimum fragment length (0), only valid with --no-spliced-alignment -X/--maxins maximum fragment length (500), only valid with --no-spliced-alignment --fr/--rf/--ff -1, -2 mates align fw/rev, rev/fw, fw/fw (--fr) --no-mixed suppress unpaired alignments for paired reads --no-discordant suppress discordant alignments for paired reads Output: -t/--time print wall-clock time taken by search phases --un write unpaired reads that didn't align to --al write unpaired reads that aligned at least once to --un-conc write pairs that didn't align concordantly to --al-conc write pairs that aligned concordantly at least once to (Note: for --un, --al, --un-conc, or --al-conc, add '-gz' to the option name, e.g. --un-gz , to gzip compress output, or add '-bz2' to bzip2 compress output.) --summary-file print alignment summary to this file. --new-summary print alignment summary in a new style, which is more machine-friendly. --quiet print nothing to stderr except serious errors --met-file send metrics to file at (off) --met-stderr send metrics to stderr (off) --met report internal counters & metrics every secs (1) --no-head supppress header lines, i.e. lines starting with @ --no-sq supppress @SQ header lines --rg-id set read group id, reflected in @RG line and RG:Z: opt field --rg add ("lab:value") to @RG line of SAM header. Note: @RG line only printed when --rg-id is set. --omit-sec-seq put '*' in SEQ and QUAL fields for secondary alignments. Performance: -o/--offrate override offrate of index; must be >= index's offrate -p/--threads number of alignment threads to launch (1) --reorder force SAM output order to match order of input reads --mm use memory-mapped I/O for index; many 'hisat2's can share Other: --qc-filter filter out reads that are bad according to QSEQ filter --seed seed for random number generator (0) --non-deterministic seed rand. gen. arbitrarily instead of using read attributes --remove-chrname remove 'chr' from reference names in alignment --add-chrname add 'chr' to reference names in alignment --version print version information and quit -h/--help print this usage message ---------------------------------------------- Program options for hisat2-build: HISAT2 version 2.1.0 by Daehwan Kim (infphilo@gmail.com, http://www.ccb.jhu.edu/people/infphilo) Usage: hisat2-build [options]* reference_in comma-separated list of files with ref sequences hisat2_index_base write ht2 data to files with this dir/basename Options: -c reference sequences given on cmd line (as ) --large-index force generated index to be 'large', even if ref has fewer than 4 billion nucleotides -a/--noauto disable automatic -p/--bmax/--dcv memory-fitting -p number of threads --bmax max bucket sz for blockwise suffix-array builder --bmaxdivn max bucket sz as divisor of ref len (default: 4) --dcv diff-cover period for blockwise (default: 1024) --nodc disable diff-cover (algorithm becomes quadratic) -r/--noref don't build .3/.4.ht2 (packed reference) portion -3/--justref just build .3/.4.ht2 (packed reference) portion -o/--offrate SA is sampled every 2^offRate BWT chars (default: 5) -t/--ftabchars # of chars consumed in initial lookup (default: 10) --localoffrate SA (local) is sampled every 2^offRate BWT chars (default: 3) --localftabchars # of chars consumed in initial lookup in a local index (default: 6) --snp SNP file name --haplotype haplotype file name --ss Splice site file name --exon Exon file name --seed seed for random number generator -q/--quiet verbose output (for debugging) -h/--help print detailed description of tool and its options --usage print this usage message --version print version information and quit ---------------------------------------------- Program options for samtools_sort: samtools sort: failed to read header from "-" sort: invalid option -- 'h' Usage: samtools sort [options...] [in.bam] Options: -l INT Set compression level, from 0 (uncompressed) to 9 (best) -m INT Set maximum memory per thread; suffix K/M/G recognized [768M] -n Sort by read name -t TAG Sort by value of TAG. Uses position as secondary index (or read name if -n is set) -o FILE Write final output to FILE rather than standard output -T PREFIX Write temporary files to PREFIX.nnnn.bam --no-PG do not add a PG line --input-fmt-option OPT[=VAL] Specify a single input file format option in the form of OPTION or OPTION=VALUE -O, --output-fmt FORMAT[,OPT[=VAL]]... Specify output format (SAM, BAM, CRAM) --output-fmt-option OPT[=VAL] Specify a single output file format option in the form of OPTION or OPTION=VALUE --reference FILE Reference sequence FASTA FILE [null] -@, --threads INT Number of additional threads to use [0] --verbosity INT Set level of verbosity ---------------------------------------------- Program options for samtools_view: [main_samview] fail to read the header from "-". [main_samview] fail to read the header from "-". ---------------------------------------------- Program options for samtools_index: Usage: samtools index [-bc] [-m INT] [out.index] Options: -b Generate BAI-format index for BAM files [default] -c Generate CSI-format index for BAM files -m INT Set minimum interval size for CSI indices to 2^INT [14] -@ INT Sets the number of threads [none] index: invalid option -- 'h' Usage: samtools index [-bc] [-m INT] [out.index] Options: -b Generate BAI-format index for BAM files [default] -c Generate CSI-format index for BAM files -m INT Set minimum interval size for CSI indices to 2^INT [14] -@ INT Sets the number of threads [none] ----------------------------------------------