Installing FastQC ------------------- FastQC is a java application. In order to run it needs your system to have a suitable Java Runtime Environment (JRE) installed. Before you try to run FastQC you should therefore ensure that you have a suitable JRE. There are a number of different JREs available however the one we have tested is the v1.6 JRE from Oracle. This is available for a number of different platforms from www.java.com (click the download now button at the top). If you're not sure whether you have java installed then you can test this from a command prompt. To get a command prompt try: Windows: Select Start > Run, and type 'cmd' (no quotes) in the box which appears, press OK MaxOSX: Run Applications > Utilities > Terminal Linux: From your applications menu look for an application called 'Terminal' or 'Konsole'. Either of these will give you a usable shell. At the command prompt type 'java -version' and press enter. You should see something like: java version "1.6.0_17" Java(TM) SE Runtime Environment (build 1.6.0_17-b04-248-10M3025) Java HotSpot(TM) Client VM (build 14.3-b01-101, mixed mode) If you get an error then you don't have java installed. If the version listed on the first line is less than 1.5 then you might have problems running FastQC. Actually installing FastQC is as simple as unzipping the zip file it comes in into a suitable location. That's it. Once unzipped it's ready to go. Running FastQC -------------- You can run FastQC in one of two modes, either as an interactive graphical application in which you can dynamically load FastQ files and view their results. Alternatively you can run FastQC in a non-interactive mode where you specify the files you want to process on the command line and FastQC will generate an HTML report for each file without launching a user interface. This would allow FastQC to be run as part of an analysis pipeline. Running FastQC Interactively ---------------------------- Windows: Simply double click on the run_fastqc bat file. If you want to make a pretty shortcut then we've included an icon file in the top level directory so you don't have to use the generic bat file icon. MacOSX: There is an application bundle for MacOSX which you can use to install and run FastQC. Just drag the application from the disk image to your Applications folder (or wherever you want to install the program). Linux: We have included a wrapper script, called 'fastqc' which is the easiest way to start the program. The wrapper is in the top level of the FastQC installation. You may need to make this file executable: chmod 755 fastqc ..but once you have done that you can run it directly ./fastqc ..or place a link in /usr/local/bin to be able to run the program from any location: sudo ln -s /path/to/FastQC/fastqc /usr/local/bin/fastqc Running FastQC as part of a pipeline ------------------------------------ To run FastQC non-interactively you should use the fastqc wrapper script to launch the program. You will probably want to use the zipped install file on every platform (even OSX). To run non-interactively you simply have to specify a list of files to process on the commandline fastqc somefile.txt someotherfile.txt You can specify as many files to process in a single run as you like. If you don't specify any files to process the program will try to open the interactive application which may result in an error if you're running in a non-graphical environment. There are a few extra options you can specify when running non-interactively. Full details of these can be found by running fastqc --help By default, in non-interactive mode FastQC will create an HTML report with embedded graphs, but also a zip file containing individual graph files and additional data files containing the raw data from which plots were drawn. The zip file will not be extracted by default but you can enable this by adding: --extract To the launch command. If you want to save your reports in a folder other than the folder which contained your original FastQ files then you can specify an alternative location by setting a --outdir value: --outdir=/some/other/dir/ If you want to run fastqc on a stream of data to be read from standard input then you can do this by specifing 'stdin' as the name of the file to be processed and then streaming uncompressed fastq format data to the program. For example: zcat *fastq.gz | fastqc stdin Customising the report output ----------------------------- If you want to run FastQC as part of a sequencing pipeline you may wish to change the formatting of the report to add in your own branding or to include extra information. In the Templates directory you will find a file called 'header_template.html' which you can edit to change the look of the report. This file contains all of the header for the report file, including the CSS section and you can alter this however you see fit. Whilst you can make whatever changes you like you should probably leave in place the
structure of the html template since later code will expect to close the main div which is left open at the end of the header. There is no facility to change the code in the main body of the report or the footer (although you can of course change the styling). The text tags @@FILENAME@@ and @@DATE@@ are placeholders which are filled in when the report it created. You can use these placeholders in other parts of the header if you wish.