snap-aligner_paired - scalable nucleotide alignment program
Welcome to SNAP version 1.0.0.
Too few parameters Usage: snap-aligner paired <index-dir>
<inputFile(s)> [<options>] where <input file(s)> is a list
of files to process.
- -o
- filename output alignments to filename in SAM or BAM format, depending on
the file extension or explicit type specifier (see below). Use a dash with
an explicit type specifier to write to stdout, so for example -o
-sam - would write SAM output to stdout
- -d
- maximum edit distance allowed per read or pair (default: 27)
- -n
- number of seeds to use per read
- -sc
- Seed coverage (i.e., readSize/seedSize). Floating point. Exclusive with
-n. (default uses -n)
- -h
- maximum hits to consider per seed (default: 300)
- -ms
- minimum seed matches per location (default: 1)
- -t
- number of threads (default is one per core)
- -b-
- Don't bind each thread to its processor (note the double dash)
- -P
- disables cache prefetching in the genome; may be helpful for machines with
small caches or lots of cores/cache
- -so
- sort output file by alignment location
- -sm
- memory to use for sorting in Gbytes. Default is 1 Gbyte/thread.
- -sid
- Specifies the sort intermediate directory. When SNAP is sorting, it aligns
the reads in the order in which they come in, and writes the aligned reads
in batches to a temporary file. When the aligning is done, it does a merge
sort from the temporary file into the final output file. By default, the
intermediate file is in the same directory as the output file, but for
performance or space reasons, you might want to put it elsewhere. If so,
use this option.
- -x
- explore some hits of overly popular seeds (useful for filtering)
- -S
- suppress additional processing (sorted BAM output only) i=index,
d=duplicate marking
- -f
- stop on first match within edit distance limit (filtering mode)
- -F
- filter output (a=aligned only, s=single hit only (MAPQ >= 10),
u=unaligned only, l=long enough to align (see -mrl))
- -E
- an alternate (and fully general) way to specify filter options. Emit only
these types s = single hit (MAPQ >= 10), m = multiple hit (MAPQ <
10), x = not long enough to align, u = unaligned, b = filter must apply to
both ends of a paired-end read. Combine the letters after -E, so
for example -E smu will emit all reads that aren't too short/have
too many Ns (because it leaves off l). -E smx is the same as
-F a, -E ux is the same as -F u, and so forth. When
filtering in paired-end mode (either with -F or -E) unless
you specify the b flag a read will be emitted if it's mate pair passes the
filter Even if the read itself does not. If you specify b mode, then a
read will be emitted only if it and its partner both pass the filter.
- -I
- ignore IDs that don't match in the paired-end aligner
-Cxx must be followed by two + or - symbols saying
whether to clip low-quality
- bases from front and back of read respectively; default: back only
(-C-+)
- -=
- use the new style CIGAR strings with = and X rather than M. The opposite
of -M
- -pf
- specify the name of a file to contain the run speed
- -hp
- Indicates to use huge pages (this may speed up alignment and slow down
index load).
- -D
- Specifies the extra search depth (the edit distance beyond the best hit
that SNAP uses to compute MAPQ). Default 1
- -rg
- Specify the default read group if it is not specified in the input
file
- -R
- Specify the entire read group line for the SAM/BAM output. This must
include an ID tag. If it doesn't start with '@RG' SNAP will add that.
Specify tabs by \t. Two backslashes will generate a single backslash.
backslash followed by anything else is illegal. So, '-R @RG\tID:foo\tDS:my
data' would generate reads with default tag foo, and an @RG line that also
included the DS:my data field.
- -sa
- Include reads from SAM or BAM files with the secondary (0x100) or
supplementary (0x800) flag set; default is to drop them.
- -om
- Output multiple alignments. Takes as a parameter the maximum extra edit
distance relative to the best alignment to allow for secondary
alignments
- -omax Limit the number
of alignments per read generated by -om.
- This means that if -om would generate more
- than -omax secondary alignments, SNAP will write out only the best
-omax of them, where 'best' means 'with the lowest edit distance'.
Ties are broken arbitrarily.
-mpc Limit the number of alignments generated by
-om to this many per contig (chromosome/FASTA entry);
- 'mpc' means 'max per contig; default unlimited.
- This filter is applied prior to -omax. The primary alignment
- is counted.
- -pc
- Preserve the soft clipping for reads coming from SAM or BAM files
- -xf
- Increase expansion factor for BAM and GZ files (default 1.0)
-hdp Use Hadoop-style prefixes (reporter:status:...) on
error messages, and emit hadoop-style progress messages
- -mrl Specify the minimum
read length to align, reads shorter than this (after clipping) stay
unaligned.
- This should be
- a good bit bigger than the seed
length or you might get some questionable alignments.
- Default 50
- -map Use file mapping to
load the index rather than reading it.
- This might speed up index loading in cases
- where SNAP is run repatedly on the same index, and the index is larger
than half of the memory size of the machine. On some operating systems,
loading an index with -map is much slower than without if the index
is not in memory. You might consider adding -pre to prefetch the
index into system cache when loading with -map when you don't
expect the index to be in cache. This is the default
-map- Do not map the index, read it using standard
read/write calls.
- -pre Prefetch the index
into system cache.
- This is only meaningful with -map, and only helps if the index is
not
- already in memory and your operating system is slow at reading mapped
files (i.e., some versions of Linux, but not Windows). This is the default
on Linux.
- -pre- Do not prefetch
the index into system cache.
- This is the default on Windows.
- -lp
- Run SNAP at low scheduling priority (Only implemented on Windows)
- -nu
- No Ukkonen: don't reduce edit distance search based on prior candidates.
This option is purely for evaluating the performance effect of using
Ukkonen's algorithm rather than Smith-Waterman, and specifying it will
slow down execution without improving the alignments.
- -no
- No Ordering: don't order the evalutation of potential alignments so as to
select more likely candidates first. This option is purely for evaluating
the performance effect of the read evaluation order, and specifying it
will slow down execution without improving alignments.
- -nt
- Don't truncate searches based on missed seed hits. This option is purely
for evaluating the performance effect of candidate truncation, and
specifying it will slow down execution without improving alignments.
- -wbs
- Write buffer size in megabytes. Don't specify this unless you've gotten an
error message saying to make it bigger. Default 16.
- -di
- Drop the index after aligning and before sorting. This frees up memory for
the sort at the expense of not having the index loaded for your next
run.
- -kts
- Kill if too slow. Monitor our progress and kill ourself if we're not
moving fast enough. This is intended for use on machines with limited
memory, where some alignment tasks will push SNAP into paging, and take
disproportinaltely long. This allows the script to move on to the next
alignment. Only works when generating output, and not during the sort
phase. If you're running out of memory sorting, try using -di.
- -pro
- Profile alignment to give you an idea of how much time is spent aligning
and how much waiting for IO
-proAg Profile affine-gap scoring to show how often it
forces single-end alignment
- -ae
- Apply the end-of-contig soft clipping before the -om processing
rather than after it. A read that's soft clipped because of hanging off
one end or the other of a contig does not have a penalty in its NM tag,
but it does in SNAP's internal scoring. This flag says to use the NM value
for -om processing rather than SNAP's internal score.
- -is
- Write SNAP's internal score for an alignment into the output. The value
following -is specifies the tag to use, and must be a two letter
value starting with X, Y or Z. So, -is ZQ will cause SNAP to write
ZQ:i:3 on a read with internal score 3. Generally, the internal scores are
the same as the NM values, except that they contain penalties for soft
clipping reads that hang over the end of contigs (but not for soft
clipping that's due to # quality scores or that was present in the input
SAM/BAM file and retained due to -pc)
- -G-
- disable affine gap scoring (default: true) Scoring parameters (works only
when -G- is not used)
- cost for match -gm (default: 1) cost for substitution -gs
(default: 4) cost for opening a gap -go (default: 6) cost for
extending a gap -ge (default: 1)
- -A-
- Disable ALT awareness. The default is to try to map reads to the primary
assembly and only to choose ALT alignments when they're much better, and
to compute MAPQ for non-ALT alignments using only non-ALT hits. This flag
disables that behavior and results in ALT-oblivious behavior.
- -ea
- Emit ALT alignments. When the aligner is ALT aware (i.e., -A- isn't
specified) if it finds an ALT alignment that would have been the primary
alignment if -A- had been specified but isn't without -A-,
SNAP will emit the read with the supplementary alignment flag set and MAPQ
computed across all potential mappings, both primary and ALT
- -asg Maximum score gap to
prefer a non-ALT alignment.
- If the best non-ALT alignment is more than this much worse than the best
ALT alignment
- emit the ALT alignment as the primary result rather than as a
supplementary result. (default: 16)
You may process more than one alignment without restarting SNAP,
and if possible without reloading the index. In order to do this, list on
the command line all of the parameters for the first alignment, followed by
a comma (separated by a space from the other parameters) followed by the
parameters for the next alignment (including single or paired). You may have
as many of these as you please. If two consecutive alignments use the same
index, it will not be reloaded. So, for example, you could do 'snap-aligner
single hg19-20 foo.fq -o foo.sam , paired hg19-20 end1.fq end2.fq
-o paired.sam' and it would not reload the index between the single
and paired alignments. When specifying an input or output file, you can
simply list the filename, in which case SNAP will infer the type of the file
from the file extension (.sam or .bam for example), or you can explicitly
specify the file type by preceding the filename with one of the
- following type specifiers (which are case sensitive):
-fastq
-compressedFastq
-sam
-bam
-pairedFastq
-pairedInterleavedFastq
-pairedCompressedInterleavedFastq
So, for example, you could specify -bam input.file to make
SNAP treat input.file as a BAM file, even though it would ordinarily assume
a FASTQ file for input or a SAM file for output when it doesn't recoginize
the file extension. In order to use a file name that begins with a '-' and
not have SNAP treat it as a switch, you must explicitly specify the type.
But really, that's just confusing and you shouldn't do it. Input and output
may also be from/to stdin/stdout. To do that, use a - for the input or
output file name and give an explicit type specifier. So, for example,
snap-aligner single myIndex -fastq - -o -sam - would
read FASTQ from stdin and write SAM to stdout.
- -s
- min and max spacing to allow between paired ends (default: 0 1000). If it
can't find an alignment in that range, it will run both reads through the
single-end aligner.
-ins Infer inter-read spacing by periodially looking at
the observed distances
- -fs
- force spacing to lie between min and max.
- -H
- max hits for intersecting aligner (default: 4000).
-mcp specifies the maximum candidate pool size (An
internal data structure.
- Only increase this if you get an error message saying to do so. If you're
running out of memory, you may want to reduce it. Default: 1000000)
-F b additional option to -F to require both
mates to satisfy filter (default is just one)
- If you specify -F b together with one of the other -F
options, -F b MUST be second
- -ku
- Keep unpaired-looking reads in SAM/BAM input. Ordinarily, if a read
doesn't specify mate information (RNEXT field is * and/or PNEXT is 0) then
the code that matches reads will immdeiately discard it. Specifying this
flag may cause large memory usage for some input files, but may be
necessary for some strangely formatted input files. You'll also need to
specify this flag for SAM/BAM files that were aligned by a single-end
aligner.
- -N
- max seeds when falling back to the single-end mode when doing paired-end.
Default: 25
- -en
- min edit distance for a read aligned as non-ALT by the paired-end aligner
to be reconsidered for a better alignment by the single-end aligner.
Default: 3
- -es
- min total edit distance by which a read pair aligned as ALT needs to be
better than non-ALT alignments to skip single-end realignment. Default:
3
- -eg
- min affine gap score improvement needed for single-end alignments to be
considered over paired-end alignments. Default: 15