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BAM is the compressed binary version of the Sequence Alignment/Map (SAM) format, a compact and index-able representation of nucleotide sequence alignments. Many next-generation sequencing and analysis tools work with SAM/BAM. For custom track display, the main advantage of indexed BAM over PSL and other human-readable alignment formats is that only the portions of the files needed to display a particular region are transferred to UCSC. This makes it possible to display alignments from files that are so large that the connection to UCSC would time out when attempting to upload the whole file to UCSC. Both the BAM file and its associated index file remain on your web-accessible server (http, https, or ftp), not on the UCSC server. UCSC temporarily caches the accessed portions of the files to speed up interactive display. If you do not have access to a web-accessible server and need hosting space for your BAM files, please see the Hosting section of the Track Hub Help documentation.
The typical workflow for generating a BAM custom track is this:
samtools
with
no command line arguments; it should print a brief usage message. For help with
samtools
, please contact the SAM tools mailing list. samtools view -S -b -o my.bam my.sam
If converting a SAM file that does not have a proper header, the -t
or
-T
option is necessary. For more information about the command, run
samtools view
with no other arguments. samtools sort my.bam my.sorted
samtools index my.sorted.bam
The sort command appends .bam
to my.sorted
, creating a BAM file of
alignments ordered by leftmost position on the reference assembly. The index command generates a
new file, my.sorted.bam.bai
, with which genomic coordinates can quickly be translated
into file offsets in my.sorted.bam
.my.sorted.bam
and
my.sorted.bam.bai
) to an http, https, or ftp location. Note that the Genome Browser
looks for an index file with the same URL as the BAM file with the .bai suffix added. If your
hosting site does not use the filename as the URL link, you will have to specifically
call the location of this .bam.bai index file with the bigDataIndex
keyword.
This keyword is relevant in Custom Tracks
and Track Hubs. You can read more about bigDataIndex in
the TrackDb Database Definition page. track type=bam name="My BAM" bigDataUrl=http://myorg.edu/mylab/my.sorted.bam
Again, in addition to http://myorg.edu/mylab/my.sorted.bam, the associated index file
http://myorg.edu/mylab/my.sorted.bam.bai must also be available at the same
location. If not, you can specify the URL with the
bigDataUrl=http://myorg.edu/mylab/my.sorted.bam.bai
All options are placed in a single line separated by spaces. In the example below, the lines are broken only for readability. If you copy/paste this example, you must remove the line breaks. Click here for a text version that you can paste without editing.
track type=bam bigDataUrl=http://...
pairEndsByName=.
pairSearchRange=N
bamColorMode=strand|gray|tag|off
bamGrayMode=aliQual|baseQual|unpaired
bamColorTag=XX
minAliQual=N
showNames=on|off
name=track_label
description=center_label
visibility=display_mode
priority=priority
db=db
maxWindowToDraw=N
chromosomes=chr1,chr2,...
The track type and bigDataUrl are REQUIRED:
type=bam bigDataUrl=http://myorg.edu/mylab/my.sorted.bam
The remaining settings are OPTIONAL. Some are specific to BAM:
pairEndsByName any value # presence indicates paired-end alignments
pairSearchRange N # max distance between paired alignments, default 20,000 bases
bamColorMode strand|gray|tag|off # coloring method, default is strand
bamGrayMode aliQual|baseQual|unpaired # grayscale metric, default is aliQual
bamColorTag XX # optional tag for RGB color, default is "YC"
minAliQual N # display only items with alignment quality at least N, default 0
showNames on|off # if off, don't display query names, default is on
Other optional settings are not specific to BAM, but relevant:
name track label # default is "User Track"
description center label # default is "User Supplied Track"
visibility squish|pack|full|dense|hide # default is hide (will also take numeric values 4|3|2|1|0)
bigDataUrl https://your.bam.bai.com # default is the bigDataUrl with .bai added to the end
priority N # default is 100
db genome database # e.g. hg18 for Human Mar. 2006
maxWindowToDraw N # don't display track when viewing more than N bases
chromosomes chr1,chr2,... # track contains data only on listed reference assembly sequences
doWiggle on|off # if on, show data as density graph, default is off
The BAM track configuration help page describes
the BAM track configuration page options corresponding to pairEndsByName
,
minAliQual
, bamColorMode
, bamGrayMode
and
bamColorTag
in more detail.
pairSearchRange
applies only when pairEndsByName
is given. It allows for
a tradeoff of display speed vs. completeness of pairing the paired-end alignments. When paired ends
are split or separated by large gaps or introns, but one is viewing a small genomic region, it is
necessary to search a large number of bases upstream and downstream of the viewed region in order
to find mates of the alignments in the viewed region. However, searching a very large region can be
slow, especially when the alignments have deep coverage of the genome. To ensure that all properly
paired mates will be found, pairSearchRange
should be set to the largest genomic size
of a mapped pair. However, it can be set to a smaller size if necessary to speed up the display, at
the cost of some items being displayed as unpaired when the mate is too far outside the viewed
window.
In this example, you will create a custom track for an indexed BAM file that is already on a public server — alignments of sequence generated by the 1000 Genomes Project.
You can paste the URL http://genome.ucsc.edu/goldenPath/help/examples/bamExample.bam
directly into the custom track management page
for the human assembly hg18 (May 2006), then press the submit button. On the following
page, press the chr21 link in the custom track and navigate to position
chr21:33,038,946-33,039,092 to see the reads in the new BAM track.
Alternatively, you can specify more visualization options by creating a "track" line. The line breaks inserted here for readability must be removed before submitting the track line:
track type=bam name="BAM Example One" description="Bam Ex. 1: 1000 Genomes read alignments (individual NA12878)"
pairEndsByName=. pairSearchRange=10000 chromosomes=chr21 bamColorMode=gray maxWindowToDraw=200000
db=hg18 visibility=pack
bigDataUrl=http://genome.ucsc.edu/goldenPath/help/examples/bamExample.bam
Include the following "browser" line to view a small region of chromosome 21 with alignments from the .bam file:
browser position chr21:33,038,946-33,039,092
Note if you copy/paste the above example, you must remove the line breaks (or, click here for a text version that you can paste without editing).
Paste the "browser" line and "track" line into the custom track management page for the human assembly hg18 (May 2006), then press the "submit" button. On the following page, press the chr21 link in the custom track listing to view the BAM track in the Genome Browser.
In this example, you will create indexed BAM from an existing SAM file. First, save this SAM file
samExample.sam to your machine. Perform steps
1 and 3-7 in the workflow described above, but substituting samExample.sam
for
my.sam
. On the custom track management
page, click the "add custom tracks" button if necessary and make sure that the genome
is set to Human and the assembly is set to Mar. 2006 (hg18) before pasting the track line and
submitting. This track line is a little nicer than the one shown in step 6, but remember to remove
the line breaks that have been added to the track line for readability (or, click
here for a text version that you can paste without
editing):
track type=bam name="BAM Example Two"
bigDataUrl=http://myorg.edu/mylab/my.sorted.bam
description="Bam Ex. 2: Simulated RNA-seq read alignments" visibility=squish
db=hg18 chromosomes=chr21
browser position chr21:33,037,317-33,038,137
browser pack mrna
If you would like to share your BAM data track with a colleague, learn how to create a sharable URL by looking at this page.