Difference between revisions of "FASTQC and MultiQC"

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Revision as of 15:32, 6 July 2016

Introduction

Quality visualization

Help file

SYNOPSIS

   fastqc seqfile1 seqfile2 .. seqfileN

   fastqc [-o output dir] [--(no)extract] [-f fastq|bam|sam] [-c contaminant file] seqfile1 .. seqfileN

DESCRIPTION

   FastQC reads a set of sequence files and produces from each one a quality
   control report consisting of a number of different modules, each one of 
   which will help to identify a different potential type of problem in your
   data.
   
   If no files to process are specified on the command line then the program
   will start as an interactive graphical application.  If files are provided
   on the command line then the program will run with no user interaction
   required.  In this mode it is suitable for inclusion into a standardised
   analysis pipeline.
   
   The options for the program as as follows:
   
   -h --help       Print this help file and exit
   
   -v --version    Print the version of the program and exit
   
   -o --outdir     Create all output files in the specified output directory.
                   Please note that this directory must exist as the program
                   will not create it.  If this option is not set then the 
                   output file for each sequence file is created in the same
                   directory as the sequence file which was processed.
                   
   --casava        Files come from raw casava output. Files in the same sample
                   group (differing only by the group number) will be analysed
                   as a set rather than individually. Sequences with the filter
                   flag set in the header will be excluded from the analysis.
                   Files must have the same names given to them by casava
                   (including being gzipped and ending with .gz) otherwise they
                   won't be grouped together correctly.
                   
   --nofilter      If running with --casava then don't remove read flagged by
                   casava as poor quality when performing the QC analysis.
                  
   --extract       If set then the zipped output file will be uncompressed in
                   the same directory after it has been created.  By default
                   this option will be set if fastqc is run in non-interactive
                   mode.
                   
   -j --java       Provides the full path to the java binary you want to use to
                   launch fastqc. If not supplied then java is assumed to be in
                   your path.
                  
   --noextract     Do not uncompress the output file after creating it.  You
                   should set this option if you do not wish to uncompress
                   the output when running in non-interactive mode.
                   
   --nogroup       Disable grouping of bases for reads >50bp. All reports will
                   show data for every base in the read.  WARNING: Using this
                   option will cause fastqc to crash and burn if you use it on
                   really long reads, and your plots may end up a ridiculous size.
                   You have been warned!
                   
   -f --format     Bypasses the normal sequence file format detection and
                   forces the program to use the specified format.  Valid
                   formats are bam,sam,bam_mapped,sam_mapped and fastq
                   
   -t --threads    Specifies the number of files which can be processed
                   simultaneously.  Each thread will be allocated 250MB of
                   memory so you shouldn't run more threads than your
                   available memory will cope with, and not more than
                   6 threads on a 32 bit machine
                 
   -c              Specifies a non-default file which contains the list of
   --contaminants  contaminants to screen overrepresented sequences against.
                   The file must contain sets of named contaminants in the
                   form name[tab]sequence.  Lines prefixed with a hash will
                   be ignored.

   -a              Specifies a non-default file which contains the list of
   --adapters      adapter sequences which will be explicity searched against
                   the library. The file must contain sets of named adapters
                   in the form name[tab]sequence.  Lines prefixed with a hash
                   will be ignored.
                   
   -l              Specifies a non-default file which contains a set of criteria
   --limits        which will be used to determine the warn/error limits for the
                   various modules.  This file can also be used to selectively 
                   remove some modules from the output all together.  The format
                   needs to mirror the default limits.txt file found in the
                   Configuration folder.