Date: Thu, 28 Mar 2024 20:16:28 +0700 (NOVT)
Message-ID: <213130132.86676.1711631788055@ugene.net>
Subject: Exported From Confluence
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When you select the item in the main menu, the Align Short Reads dialog appears. Set value of the Align short reads method pa=
rameter to Bowtie. The dialog looks as follows:
=20
=20
=
=20
=20
There are the following parameters:
Ref=
erence sequence =E2=80=94 DNA sequence to align short reads to. T=
his parameter is required.
Res=
ult file name =E2=80=94 file in SAM format to write the result of=
the alignment into. This parameter is required.
Library - si=
ngle-end or paired-end reads.
Pre=
built index =E2=80=94 check this box to use an index file instead=
of a source reference sequence. The index is a set of 6 files with suffixe=
s .1.ebwt, .2.ebwt, .3.ebwt, .4.ebwt, .rev.1.ebwt, and .rev.2.ebwt. The ind=
ex is created during the alignment. Also you can build it manually=
em>.
SAM=
output =E2=80=94 always save the output file in the SAM format (=
the option is disabled for Bowtie).
Sho=
rt reads =E2=80=94 each added short read is a small DNA sequence =
file. At least one read should be added.
You can also configure other parameters. =
They are the same as in the original Bowtie (you can read detailed description of the parameters on the Bowtie manual page).
Select one of the following alignment mod=
es:
The -n alignment mode:
When the -n mode is selected, =
Bowtie determines which alignments are valid according to the fol=
lowing policy. Alignments may have no more than N mismatches (where N is a =
number 0-3) in the first L bases (where L is a number 5 or greater, set wit=
h Seed length) on the high-quality (left) =
end of the read. The sum of the Phred quality values at all mismatched posi=
tions (not just in the seed) may not exceed E (set with Maq error). Where qualities are unavailable (e.g. if the read=
s are from a FASTA file), the Phred quality defaults to 40.
The -v alignment mode:
In -v mode, alignments may have no more than V mismatches, wher=
e V may be a number from 0 through 3. Quality values are ignored. The =
-v mode is mutually exclusive with the&nbs=
p;-n mode.
The following parameters are available:=
p>
Maq error (=E2=80=93maqerr) =E2=80=94 maximum permitted total o=
f quality values at all mismatched read positions throughout the entire ali=
gnment, not just in the =E2=80=9Cseed=E2=80=9D. The default is 70. By defau=
lt, Bowtie rounds quality values to t=
he nearest 10 and saturates at 30. Note that the rounding can be disabled w=
ith No Maq rounding.
Seed Length (=E2=80=93seedlen) =E2=80=94 the number of bases on=
the high-quality end of the read to which the =
-n applies. The lowest permitted setting is 5 and the default is =
28.
Maximum of backtracks (-maxbts) =E2=80=94 the maximum number of=
backtracks (default: 125 without Best, 80=
0 with Best). A =E2=80=9Cbacktrack=E2=80=
=9D is the introduction of a speculative substitution into the alignment.=
p>
Descriptors memory usage (=E2=80=93chunkmbs) =E2=80=94 the numb=
er of megabytes of memory a given thread is given to store path descriptors=
in the Best flag. Default: 64. This =
parameter is available if the Best fl=
ag is checked.
Seed (=E2=80=93seed) =E2=80=94 pseudo-random number generator.<=
/p>
The following flags are available:
Colorspace (=E2=80=93color) =E2=80=94 the input is read in colo=
rspace, colors are encoded as characters A/C/G/T (A=3Dblue, C=3Dgreen, G=3D=
orange, T=3Dred).
No Maq rounding (=E2=80=93nomaqround) =E2=80=94 Maq (Mapping an=
d Assembly with Quality) accepts quality values in the Phred quality scale,=
but internally rounds values to the nearest 10, with a maximum of 30. By d=
efault, Bowtie also rounds this way.&=
nbsp;No Maq rounding prevents this roundin=
g in Bowtie.
No forward orientation (=E2=80=93nofw) =E2=80=94 do not attempt=
to align against the forward reference strand.
No reverse-complement orientation (=E2=80=93norc) =E2=80=94 do =
not attempt to align against the reverse-complement reference strand.
Try as hard (=E2=80=93tryhard) =E2=80=94 try as hard as possibl=
e to find valid alignments when they exist, including paired-end alignments=
.
Best alignments (=E2=80=93best) =E2=80=94 make Bowtie guarantee that reported singleton alignments=
are =E2=80=9Cbest=E2=80=9D in terms of stratum (i.e. number of mismatches,=
or mismatches in the seed for the case of -n m=
ode) and in terms of the quality values at the mismatched position(s).=
All alignments (=E2=80=93all) =E2=80=94 report all valid alignm=
ents per read or pair. Validity of alignments is determined by the alignmen=
t policy (combined effects of -n mode,&nbs=
p;-v mode, Seed len=
gth, and Maq error).
Select the required parameters and press =
the Start button.
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