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使用hisat2+stringtie进行基因预测

使用hisat2+stringtie进行基因预测

作者: 花生学生信 | 来源:发表于2022-03-07 18:37 被阅读0次

    比对到转录本,预测基因

    cat id1|while read id
    do
    #hisat2-build 06/${id}.arrow.polish.fasta.masked index/${id}
    #hisat2 --dta -p 20 -x index/${id} -1 /public/home/lianglunping/work/HXM/RNA/input/HA798-normal-1A_1.fq.gz -2 /public/home/lianglunping/work/HXM/RNA/input/HA798-normal-1A_2.fq.gz -S 08sam/${id}.1.sam
    #hisat2 --dta -p 20 -x index/${id} -1 /public/home/lianglunping/work/HXM/RNA/input/HA798-roll-1A_1.fq.gz -2 /public/home/lianglunping/work/HXM/RNA/input/HA798-roll-1A_2.fq.gz -S 08sam/${id}.2.sam
    #hisat2 --dta -p 20 -x index/${id} -1 /public/home/lianglunping/work/HXM/RNA/input/NPB-1A_1.fq.gz -2 /public/home/lianglunping/work/HXM/RNA/input/NPB-1A_2.fq.gz  -S 08sam/${id}.3.sam
    #hisat2 --dta -p 20 -x index/${id} -1 /public/home/lianglunping/work/HXM/RNA/input/TR-grass-1A_1.fq.gz -2 /public/home/lianglunping/work/HXM/RNA/input/TR-grass-1A_2.fq.gz  -S 08sam/${id}.4.sam
    #hisat2 --dta -p 20 -x index/${id} -1 /public/home/lianglunping/work/HXM/RNA/input/TR-normal-1A_1.fq.gz -2 /public/home/lianglunping/work/HXM/RNA/input/TR-normal-2A_1.fq.gz  -S 08sam/${id}.5.sam
    #samtools view -bS 08sam/$id.1.sam -o 09bam/$id.1.bam
    #samtools view -bS 08sam/$id.2.sam -o 09bam/$id.2.bam
    #samtools view -bS 08sam/$id.3.sam -o 09bam/$id.3.bam
    #samtools view -bS 08sam/$id.4.sam -o 09bam/$id.4.bam
    #samtools view -bS 08sam/$id.5.sam -o 09bam/$id.5.bam
    #samtools merge -@ 88   $id.merge    09bam/$id.1.bam 09bam/$id.2.bam 09bam/$id.3.bam 09bam/$id.4.bam 09bam/$id.5.bam 
    #samtools sort -@ 88 09bam/$id.merge  -o 09bam/$id.s.bam
    stringtie -p 10 -o 11sbam/$id.gtf 09bam/$id.s.bam
    done
    

    使用TransDecoder将结果整理成evm输入格式

    /public/home/fengting/tools/TransDecoder-v5.5.0/util/gtf_genome_to_cdna_fasta.pl H7L1.gtf /public/home/fengting/task/222anno/06/H7L1.arrow.polish.fasta.masked > transcripts.fasta
    /public/home/fengting/tools/TransDecoder-v5.5.0/util/gtf_to_alignment_gff3.pl H7L1.gtf > transcripts.gff3
    TransDecoder.LongOrfs -t transcripts.fasta
    TransDecoder.Predict -t transcripts.fasta
    /public/home/fengting/tools/TransDecoder-v5.5.0/util/cdna_alignment_orf_to_genome_orf.pl \
         transcripts.fasta.transdecoder.gff3 \
         transcripts.gff3 \
         transcripts.fasta > transcripts.fasta.transdecoder.genome.gff3
    

    第一步整理后:

    hisat2 -p 50 -x masked -1 /public/home/lianglunping/work/HXM/RNA/input/HA798-normal-1A_1.fq.gz -2 /public/home/lianglunping/work/HXM/RNA/input/HA798-normal-1A_2.fq.gz | samtools sort -@ 100 > 1.bam
    hisat2 -p 50 -x masked -1 /public/home/lianglunping/work/HXM/RNA/input/HA798-roll-1A_1.fq.gz -2 /public/home/lianglunping/work/HXM/RNA/input/HA798-roll-1A_2.fq.gz | samtools sort -@ 100 > 2.bam
    hisat2 -p 50 -x masked -1 /public/home/lianglunping/work/HXM/RNA/input/NPB-1A_1.fq.gz -2 /public/home/lianglunping/work/HXM/RNA/input/NPB-1A_2.fq.gz | samtools sort -@ 100 > 3.bam
    hisat2 -p 50 -x masked -1 /public/home/lianglunping/work/HXM/RNA/input/TR-grass-1A_1.fq.gz -2 /public/home/lianglunping/work/HXM/RNA/input/TR-grass-1A_2.fq.gz | samtools sort -@ 100 > 4.bam
    hisat2 -p 50 -x masked -1 /public/home/lianglunping/work/HXM/RNA/input/TR-normal-1A_1.fq.gz -2 /public/home/lianglunping/work/HXM/RNA/input/TR-normal-2A_1.fq.gz | samtools sort -@ 100 > 5.bam
    
    samtools merge -@ 88   I.merge   1.bam 2.bam 3.bam 4.bam 5.bam 
    samtools sort -@ 88 I.merge  -o I.s.bam
    
    stringtie -p 100 -o I.gtf I.s.bam
    

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