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                                    ____ _              v0.52
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                                                round is good

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Circos - circular genome data and annotation image generator

v0.52

Martin Krzywinski
Canada's Michael Smith Genome Sciences Center
British Columbia Cancer Agency

[email protected]
mkweb.bcgsc.ca/circos

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0. INTRODUCTION
   0.a   what is circos?
   0.b   requirements

1. GETTING STARTED
   1.a   installation 
   1.b   tools

2. BUGS
   2.a 	 report bugs and comments
   2.b   known issues

3. INSTALLATION PROBLEMS
   3.a   missing modules

4. OTHER ISSUES
   4.a   configuration paths
   4.b   typical errors and how to fix them
   4.b.1 numerical parameter units

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0. INTRODUCTION

0.a  what is circos?

Circos is a program for the generation of publication-quality, 
circularly composited renditions of genomic data and related annotations. 

Circos is particularly suited for visualizing alignments, conservation and 
intra and inter-chromosomal relationships.

But wait. Also, Circos is useful to visualize any type of information that
benefits from a circular layout. Thus, although it has been designed for
the field of genomics, it is sufficiently flexible to be used in other data
domains.

0.b  requirements

Perl 5.8.x, or newer, is highly recommended. In addition to the core
modules that come with your Perl distribution, the following CPAN
modules are required

Clone
GD
GD::Polyline
List::Util
Math::Bezier
Math::BigFloat
Math::Round
Math::VecStat
Params::Validate
Readonly
Set::IntSpan (v1.11, or later)

Modules specific to Circos ship with the distribution in lib/.

Circos supports TTF fonts. A few fonts are included in fonts/.

1. GETTING STARTED

1.a  installation

> tar xvfz circos-x.xx.tgz
> cd circos-x.xx
> ./install

The install script will alter the path to the Perl binary and output
path for tutorial files. By default, the Perl in your path will be used (/bin/env perl) and /tmp will be assigned to the output directory.

If you prefer to change the paths manually, adjust the first line in
bin/circos to reflect its location. For example,

  #!/path/to/my/perl

and the dir parameter in <image> block in each tutorial/*/*/circos.conf file.

  <image>
  dir = /path/to/my/output
  ...
  </image>

Now try creating an image from one of the tutorials. 

> cd circos-x.xx
> bin/circos -conf tutorials/2/2/circos.conf

If everything goes well, the program should terminate with a line

  created image at /path/to/your/output/your-output-file.png

To get some verbose reporting, use 

> bin/circos -conf tutorials/2/2/circos.conf -debug

Please see mkweb.bcgsc.ca/circos for documentation. There are a large number
of tutorials that described how the configuration files are formatted.

1.b tools

There are several helper scripts located in tools/ that are designed to aid you in processing your data.

Many of these involve manipulating link files. These tools independent scripts and are covered in Tutorial 9.

  http://mkweb.bcgsc.ca/circos/?tutorials&id=9

The tools can be downloaded independently. Note that the stand-alone
tools distribution may contain scripts that are newer than those
bundled with Circos. To check this, look at the release date for the
archives at http://mkweb.bcgsc.ca/circos/?download

2. BUGS

2.a  report bugs and comments

I appreciate any and all comments you may have about Circos.

2.b  known issues

GD does not draw rotated text correctly when the font size is small. For example,
using a font size of 6pt, text drawn an an angle is drawn with letters upright. If you
see this, increase the font size of the text.

3. INSTALLATION PROBLEMS

3.a missing modules

In order to run Circos you may need to install some modules from CPAN (www.cpan.org). You 
will need the modules listed here

  http://mkweb.bcgsc.ca/circos/?requirements

If you run Circos and get a message like

Can't locate Config/General.pm in @INC (@INC contains: /usr/lib/perl5/5.8.0/i386-linux
-thread-multi /usr/lib/perl5/5.8.0 /usr/lib/perl5/site_perl/5.8.0/i386-linux-thread-mu
lti /usr/lib/perl5/site_perl/5.8.0 /usr/lib/perl5/site_perl /usr/lib/perl5/vendor_perl
/5.8.0/i386-linux-thread-multi /usr/lib/perl5/vendor_perl/5.8.0 /usr/lib/perl5/vendor_
perl .) at ./bin/circos line 121.

then you do not have a module installed. It may be that you have the module elsewhere,
but Perl cannot find it. In this case, the error message is barking at the fact that
Config::General is not installed.

You can install the module using CPAN (if CPAN module is installed)

> perl -MCPAN -e shell
% install Config::General

Make sure that you are using the same perl binary to install the module as for Circos.

Alternatively, you can grab the module from CPAN directly. Use search.cpan.org to find
the module.

> wget http://search.cpan.org/~tlinden/Config-General-2.31/General.pm
> tar xvfz Config-General-x.xx.tgz
> cd Config-General-x.xx.tgz
> perl Makefile.PL ; make ; make test
> make install

4. OTHER ISSUES

4.a configuration paths

If you look inside one of the configuration files you'll find
that it includes other configuration files using <<include>> and
makes relative mention of data files, such as

  file = data/5/segdup.txt

Circos tries to find the file regardless where you are running the binary from, but 
may still run into trouble finding files specified using a relative path.

To avoid problems, run circos from its distribution directory

> cd circos-x.xxx
> bin/circos -conf ...

Alternative, change all the paths in the .conf file to absolute paths. For example, from

  <<include etc/colors.conf>>

to

  <<include /path/to/your/install/circos-x.xx/etc/colors.conf>>

4.b typical errors and how to fix them

4.b.1 numerical parameter units

Many numerical parameters in the .conf files require that you specify
a unit. For a given parameter, one or more of these units may be required

  p - absolute size in pixels
  r - relative size, with the comparator being context sensitive 
      e.g. relative tick size is relative to ideogram thickness
      e.g. relative tick label offset is relative to tick size
  u - chromosome units, as defined by chromosomes_unit
  b - bases, the natural unit along the ideogram
  n - no unit, explicitly stated

If you receive an error message of the kind

The parameter [ideogram/spacing/break_style/thickness] value [0.25] does not have 
    the correct unit [saw n], which should be one of r,p at bin/circos line 3410
    main::unit_validate(0.25, 'ideogram/spacing/break_style/thickness', 'r', 'p') called atbin/circos line 3559
    main::draw_axis_break('HASH(0x8a81d10)') called at bin/circos line 872

Then you have the wrong unit. Here the parameter at fault is 

  ideogram/spacing/break_style/thickness

that is

  <ideogram>
   <spacing>
    <break_style>
     thickness = 

It is defined to be 0.25 (no unit) but requires that it have a unit of
either "r" (relative) or "p" (absolute).

As Circos matures, and the configuration file syntax stabilizes, I am
striving to standardize the requirement for units for all
parameters. Previous versions were more tolerant and attempted to
naively determine the unit automatically (e.g. if value was <1 then
the unit was assumed relative) and you may find that after upgrading
to a new version of Circos your old files are creating errors. Just
add the units and you're good to go.