FastQC的安装与使用
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FastQC的安装与使用
一、下载安装
1、准备工作---java环境
FastQC是一个java应用程序。为了运行它,需要您的系统安装合适的Java运行时环境。因此,在尝试运行FastQC之前,应该确保有一个合适的Java运行时环境。
$ java -version
openjdk version "1.8.0_192"
OpenJDK Runtime Environment (Zulu 8.33.0.1-linux64) (build 1.8.0_192-b01)
OpenJDK 64-Bit Server VM (Zulu 8.33.0.1-linux64) (build 25.192-b01, mixed mode)
2、下载FastQC
FastQC提供编译好的文件,下载后可以直接运行。也可以自己下载源代码自行编译。从网页获取下载链接,利用wget下载编译好的文件,目前最新的是v0.11.9版本。
下载网址:https://www.bioinformatics.babraham.ac.uk/projects/download.html#fastqc
# 下载
wget --no-check-certificate https://www.bioinformatics.babraham.ac.uk/projects/fastqc/fastqc_v0.11.9.zip
# 解压
unzip fastqc_v0.11.9.zip
# 添加可执行权限
cd FastQC
chmod +x fastqc
3、Conda安装
直接使用conda安装
conda install fastqc
二、软件参数和使用
1、常用的参数设置
- -h|--help: 输出帮助信息
- -o|--outdir:输出结果路径
- -t|--threads: 使用线程个数
- --extract:输出结果解压缩。
- -a|--adapters:自己指定的adapter序列。默认的序列是Illumina的接头序列,可以在FastQC/Configuration/adapter_list.txt文件中获取。
2、使用示例
$ fastqc -o ./ -t 1 Sample.R1.fastq.gz
$ ls
Sample.R1.fastq.gz Sample.R1.fastqc.zip Sample.R1.fastqc.html
下载html到本地可以用浏览器打开即可看到质控分析结果。
如果在流程中可以从Sample.R1.fastqc/fastqc_data.txt文件中读取数据。
同时输出的html的报告支持修改格式,可以替换为自己的模板。修改安装路径下的FastQC/Templates/header_template.html文件即可输出自己想要的风格。
附表:FastQC参数信息
点击查看代码
$ fastqc -h
FastQC - A high throughput sequence QC analysis tool
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.
--nano Files come from nanopore sequences and are in fast5 format. In
this mode you can pass in directories to process and the program
will take in all fast5 files within those directories and produce
a single output file from the sequences found in all files.
--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!
--min_length Sets an artificial lower limit on the length of the sequence
to be shown in the report. As long as you set this to a value
greater or equal to your longest read length then this will be
the sequence length used to create your read groups. This can
be useful for making directly comaparable statistics from datasets with somewhat variable read lengths.
-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 --contaminants Specifies a non-default file which contains the list of
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 --adapters Specifies a non-default file which contains the list of
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 --limits Specifies a non-default file which contains a set of criteria
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.
-k --kmers Specifies the length of Kmer to look for in the Kmer content
module. Specified Kmer length must be between 2 and 10. Default
length is 7 if not specified.
-q --quiet Supress all progress messages on stdout and only report errors.
-d --dir Selects a directory to be used for temporary files written when
generating report images. Defaults to system temp directory if
not specified.
BUGS
Any bugs in fastqc should be reported either to simon.andrews@babraham.ac.uk
or in www.bioinformatics.babraham.ac.uk/bugzilla/
标签:file,FastQC,will,--,fastqc,使用,output,安装 来源: https://www.cnblogs.com/Sunny-King/p/Bioinformatics-FastQC.html