## Download and install HISAT

# https://ccb.jhu.edu/software/hisat2/index.shtml

cd ~/biosoft

mkdir HISAT && cd HISAT

#### readme: https://ccb.jhu.edu/software/hisat2/manual.shtml

wget ftp://ftp.ccb.jhu.edu/pub/infphilo/hisat2/downloads/hisat2-2.0.4-Linux_x86_64.zip

unzip hisat2-2.0.4-Linux_x86_64.zip

ln -s hisat2-2.0.4 current

## ~/biosoft/HISAT/current/hisat2-build

## ~/biosoft/HISAT/current/hisat2

## Download and install StringTie

## https://ccb.jhu.edu/software/stringtie/ ## https://ccb.jhu.edu/software/stringtie/index.shtml?t=manual

cd ~/biosoft

mkdir StringTie && cd StringTie

wget http://ccb.jhu.edu/software/stringtie/dl/stringtie-1.2.3.Linux_x86_64.tar.gz

tar zxvf stringtie-1.2.3.Linux_x86_64.tar.gz

ln -s stringtie-1.2.3.Linux_x86_64 current

# ~/biosoft/StringTie/current/stringtie

while read id

do

sample=$(echo $id |cut -d" " -f 1 )

file1=$(echo $id |cut -d" " -f 2 )

file2=$(echo $id |cut -d" " -f 3 )

echo  $sample

echo $file1

echo $file2

~/biosoft/HISAT/current/hisat2  -p 4 --dta  -x  ~/reference/index/hisat/hg19/genome  -1 $file1 -2 $file2 -S $sample.hisat2.hg19.sam 2>$sample.hisat2.hg19.log &

done <$1

上面这个脚本需要一个3列的输入文件，分别是样本名，read1文件，read2文件，会产生以下的输出文件，sam文件。

while read id

do

file=$(basename $id )

sample=${file%%.*}

echo $id $sample

nohup samtools sort -@ 4 -o ${sample}.sorted.bam $id &

done <$1

最新版的samtools已经可以直接把sam文件变成排序好的bam文件啦~~~~

while read id

do

file=$(basename $id )

sample=${file%%.*}

echo $id $sample

nohup ~/biosoft/StringTie/current/stringtie  -p 4  -G ~/reference/gtf/gencode/gencode.v25lift37.annotation.gtf  -o $sample.hg19.stringtie.gtf -l $sample  $id  &

done <$1

stringTie的用法就是这样咯。没什么好讲的

 ~/biosoft/StringTie/current/stringtie   --merge -p 8 -G ~/reference/gtf/gencode/gencode.v25lift37.annotation.gtf  -o stringtie_merged.gtf  mergelist.txt

while read id

do

file=$(basename $id )

sample=${file%%.*}

echo $id $sample

nohup ~/biosoft/StringTie/current/stringtie -e -B  -G  $2  -o ballgown/$sample/$sample.hg19.stringtie.gtf   $id  &

done <$1