SUBREAD-ALIGN(1)

💖 SUBREAD-ALIGN 💖

okay bestie, welcome to subread-align(1) — the manual page for the subread-align toolkit for processing next-gen sequencing data. yes that's the manual name, it lives here now, we're keeping it. sit down, grab a boba, we're aligning reads. 🧚✨

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🌸 NAME 🌸

subread-align — toolkit for processing next-gen sequencing data 🦋

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✨ DESCRIPTION ✨

Version 2.0.4 💅

Usage:

./subread-align [options] -i <index_name> -r <input> -t <type> -o
<output>

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🔒 Mandatory arguments 🔒

these are not optional, babe. you NEED them. no cap.

• -i <string> — Base name of the index. this is the index, the whole index, and nothing but the index 💫
• -r <string> — Name of an input read file. If paired-end, this should be the first read file (typically containing "R1"in the file name) and the second should be provided via -R. Acceptable formats include gzipped FASTQ, FASTQ, gzipped FASTA and FASTA. These formats are identified automatically. so you literally don't have to tell it which one it is, it just knows 👑
• -t <int> — Type of input sequencing data. Its values include 0: RNA-seq data 1: genomic DNA-seq data.

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🎀 Optional arguments: input reads and output 🎀

the glow-up section. this is where you customize 💅

• -o <string> — Name of an output file. By default, the output is in BAM format. Omitting this option makes the output be written to STDOUT. (yes, straight to your terminal, main character energy, no file needed ✨)
• -R <string> — Name of the second read file in paired-end data (typically containing "R2" the file name).
• --SAMinput — Input reads are in SAM format.
• --BAMinput — Input reads are in BAM format.
• --SAMoutput — Save mapping results in SAM format.

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💜 Phred offset 💜

• -P <3:6> — Offset value added to the Phred quality score of each read base. '3' for phred+33 and '6' for phred+64. '3' by default.

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🔥 thresholds for mapping 🔥

• -n <int> — Number of selected subreads, 10 by default.
• -m <int> — Consensus threshold for reporting a hit (minimal number of subreads that map in consensus) . If paired-end, this gives the consensus threshold for the anchor read (anchor read receives more votes than the other read in the same pair). 3 by default
• -p <int> — Consensus threshold for the non- anchor read in a pair. 1 by default.
• -M <int> — Maximum number of mis-matched bases allowed in each reported alignment. 3 by default. Mis-matched bases found in softclipped bases are not counted. (so softclips get a free pass, iconic behaviour honestly)

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🌈 unique mapping and multi-mapping 🌈

• --multiMapping — Report multi-mapping reads in addition to uniquely mapped reads. Use "-B" to set the maximum number of equally-best alignments to be reported.
• -B <int> — Maximum number of equally-best alignments to be reported for a multi-mapping read. Equally-best alignments have the same number of mis-matched bases. 1 by default.

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🍓 indel detection 🍓

• -I <int> — Maximum length (in bp) of indels that can be detected. 5 by default. Indels of up to 200bp long can be detected. (yes, TWO HUNDRED, she's flexible 💅)
• --complexIndels — Detect multiple short indels that are in close proximity (they can be as close as 1bp apart from each other).

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🦋 read trimming 🦋

• --trim5 <int> — Trim off <int> number of bases from 5' end of each read. 0 by default.
• --trim3 <int> — Trim off <int> number of bases from 3' end of each read. 0 by default.

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💫 distance and orientation of paired end reads 💫

• -d <int> — Minimum fragment/insert length, 50bp by default.
• -D <int> — Maximum fragment/insert length, 600bp by default.
• -S <ff:fr:rf> — Orientation of first and second reads, 'fr' by default ( forward/reverse).

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👑 number of CPU threads 👑

• -T <int> — Number of CPU threads used, 1 by default.

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🌸 read group 🌸

• --rg-id <string> — Add read group ID to the output.
• --rg <string> — Add <tag:value> to the read group (RG) header in the output.

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✨ read order ✨

• --keepReadOrder — Keep order of reads in BAM output the same as that in the input file. Reads from the same pair are always placed next to each other no matter this option is specified or not. (they're clingy like that, it's cute 🫶)
• --sortReadsByCoordinates — Output location-sorted reads. This option is applicable for BAM output only. A BAI index file is also generated for each BAM file so the BAM files can be directly loaded into a genome browser. (so convenient, we love a two-for-one 💖)

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🎀 color space reads 🎀

• -b — Convert color-space read bases to base-space read bases in the mapping output. Note that read mapping is performed at color-space.

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💀 dynamic programming 💀

• --DPGapOpen <int> — Penalty for gap opening in short indel detection. -1 by default.
• --DPGapExt <int> — Penalty for gap extension in short indel detection. 0 by default.
• --DPMismatch <int> — Penalty for mismatches in short indel detection. 0 by default.
• --DPMatch <int> — Score for matched bases in short indel detection. 2 by default.

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🔥 detect structural variants 🔥

• --sv — Detect structural variants (eg. long indel, inversion, duplication and translocation) and report breakpoints. Refer to Users Guide for breakpoint reporting. (big structural drama detected, periodt)

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🌺 gene annotation 🌺

• -a — Name of an annotation file (gzipped file is accepted). GTF/GFF format by default. See -F option for more format information.
• -F — Specify format of the provided annotation file. Acceptable formats include 'GTF' (or compatible GFF format) and 'SAF'. 'GTF' by default. For SAF format, please refer to Users Guide.
• -A — Provide a chromosome name alias file to match chr names in annotation with those in the reads. This should be a twocolumn comma-delimited text file. Its first column should include chr names in the annotation and its second column should include chr names in the index. Chr names are case sensitive. No column header should be included in the file. (case sensitive, so chr1 and CHR1 are NOT the same girl, don't play 🙈)
• --gtfFeature <string> — Specify feature type in GTF annotation. 'exon' by default. Features used for read counting will be extracted from annotation using the provided value.
• --gtfAttr <string> — Specify attribute type in GTF annotation. 'gene_id' by default. Meta-features used for read counting will be extracted from annotation using the provided value.

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⭐ others ⭐

• -v — Output version of the program.

Refer to Users Manual for detailed description to the arguments.

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🫶 AUTHOR 🫶

This manpage was written by Alexandre Mestiashvili for the Debian distribution and can be used for any other usage of the program. (we stan, ty for the lore 💖)

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and that's the whole vibe check bestie — go align something iconic. 🦋✨

signing off: subread-align 2.0.3, March 2023 — SUBREAD-ALIGN(1) 💅🌸