Date: Fri, 29 Mar 2024 16:25:51 +0700 (NOVT)
Message-ID: <1068778515.89036.1711704351779@ugene.net>
Subject: Exported From Confluence
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When you select the item in the main menu, the Align Sequencing Reads dialog appear=
s. Set value of the Align short reads method parameter to BWA-MEM. The dialog looks as follo=
ws:
=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.
Pre=
built index =E2=80=94 check this box to use an index file instead=
of a source reference sequence. Also you can build it manually.<=
/p>
SAM=
output =E2=80=94 always save the output file in the SAM format (=
the option is disabled for BWA).
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.&=
nbsp;
Ind=
ex algorithm (-a) =E2=80=94 algorithm for constructing BWA i=
ndex.
It implements three d=
ifferent algorithms:
-
- is =E2=80=94 designed for short reads up to=
~200bp with low error rate (<3%). It does gapped global alignment w.r.t=
. reads, supports paired-end reads, and is one of the fastest short read al=
ignment algorithms to date while also visiting suboptimal hits.
- bwtsw =E2=80=94 is designed for long reads =
with more errors. It performs heuristic Smith-Waterman-like alignment to fi=
nd high-scoring local hits. Algorithm implemented in BWA-S=
W. On low-error short queries, BWA-SW.=
is slower and less accurate than the is&n=
bsp;algorithm, but on long reads, it is better.
- div =E2=80=94 does not work for long genome=
s.
Num=
ber of threads (-t) =E2=80=94 number of threa=
ds.
Min=
seed length (-k) =E2=80=94 minimum seed leng=
th. Matches shorter than INT will be missed=
. The alignment speed is usually insensitive to this value unless it signif=
icantly deviates 20.
Ban=
d width (-w) =E2=80=94 band width. Essentially, gaps longer=
than INT will not be found. Note that the =
maximum gap length is also affected by the scoring matrix and the hit lengt=
h, not solely determined by this option.
Dro=
poff (-d) =E2=80=94 off-diagonal X-dropoff (Z-dropoff). Sto=
p extension when the difference between the best and the current extension =
score is above |i-j|*A+INT, where i<=
span> and j are the current positions =
of the query and reference, respectively, and A&=
nbsp;is the matching score. Z-dropoff is similar to BLAST=E2=80=99s X-dropo=
ff except that it doesn=E2=80=99t penalize gaps in one of the sequences in =
the alignment. Z-dropoff not only avoids unnecessary extension, but also re=
duces poor alignments inside a long good alignment.
Internall s=
eeds length (-r) - trigger re-seeding for a MEM longer than=
minSeedLen*FLOAT. This is=
a key heuristic parameter for tuning the performance. Larger value yields =
fewer seeds, which leads to faster alignment speed but lower accuracy.
Skip seeds =
threshold (-c) - discard a MEM if it has more than INT occurence in the genome. This is an insensitive=
parameter.
Drop chain =
threshold (-D) - drop chains shorter than FLOAT fraction of the l=
ongest overlapping chain.
Rounds of m=
ate rescues (-m) - perform at most INT rounds of mate rescues for=
each read.
Skip mate r=
escue (-S) - skip mate rescue.
Skip pairin=
g (-P) - in the paired-end mode, perform SW to rescue missi=
ng hits only but do not try to find hits that fit a proper pair.
Score for a=
match (-A) - matching score.
Mismatch pe=
nalty (-B) - mismatch penalty. The sequence error rate is a=
pproximately: {.75 * exp[-log(4) * B/A]}.
Gap open penalty =
(-O) - gap open penalty. =
p>
Gap extention pen=
alty (-E) - gap extension penalty. A gap of length k costs O + k*=
E (i.e. Gap open penalty is for opening a zero-length gap).
Penalty for clipp=
ing (-L) - clipping penalty. When performing SW extension, =
BWA-MEM keeps track of the best score reaching the end of query. If this sc=
ore is larger than the best SW score minus the clipping penalty, clipping w=
ill not be applied. Note that in this case, the SAM AS tag reports the best=
SW score; clipping penalty is not deducted.
Penalty unpaired =
(-U) - penalty for an unpaired read pair. BWA-MEM scores an=
unpaired read pair as scoreRead1+scoreRead2-INT=
and scores a paired as scoreRead1+scoreRead2-insertPenalty. It compar=
es these two scores to determine whether we should force pairing.
Score threshold (=
-T) - don=E2=80=99t output alignment with score lower than =
score threshold. This option only affects output.
Select the required parameters and press =
the Start button.
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