对测序数据进行质控时,通常考虑三点:
1.quality trimming
2.adapter removal
3.contaminant filtering
除杂时我们用到了sickle,它能去除低质量的reads还能保证双端序列的完整性
下载地址:
https://github.com/najoshi/sickle
安装步骤:
cd ~/biosoft mkdir sickle && cd sickle #建立文件夹
wget https://codeload.github.com/najoshi/sickle/zip/master -O sickle.zip #进行下载
unzip sickle.zip #解压
cd sickle-master
make #编译
~/biosoft/sickle/sickle-master/sickle -h #查看帮助
命令语句:
~/biosoft/sickle/sickle-master/sickle pe -f test1.fastq -r test2.fastq -t sanger -o trimmed_output_file1.fastq -p trimmed_output_file2.fastq
使用参数:
pe : use paired-end mode
-f training/rnaseq/ERR022486_chr22_read1.fastq.gz : the fastq file for read 1
-r training/rnaseq/ERR022486_chr22_read2.fastq.gz : the fastq file for read 2
-t sanger : the quality encoding. All data downloaded from EBI or NCBI will be "sanger" encoded. For an explanation:<http://en.wikipedia.org/wiki/FASTQ_format#Encoding>
-o ERR022486_chr22_read1_trim.fastq : the output file for trimmed reads from read 1
-p ERR022486_chr22_read2_trim.fastq : the output file for trimmed reads from read 2
-s ERR022486_chr22_single_trim.fastq : the output file for reads where the mate has failed the quality or length filter
-q 30 : the quality value to use. Bases below this will be trimmed, using a sliding window
-l 15 : the minimum length allowed after trimming. Here we remove reads with less than 15bp
给出的结果格式: