lumpy 结构突变分析流程

Reply to lumpy 结构突变分析流程 on Mon, 25 Oct 2021 02:38:40 GMT

anneng — Mon, 25 Oct 2021 02:38:40 GMT

svtools vcfpaste
-m merged.vcf
-f cn.list
-q
| bgzip -c \

merged.sv.gt.cn.vcf.gz

IOError: [Errno 24] Too many open files: '../cn/ZGSP-9.vcf'
ulimit -n 2048 扩大文件限制

Reply to lumpy 结构突变分析流程 on Sun, 24 Oct 2021 04:02:22 GMT

anneng — Sun, 24 Oct 2021 04:02:22 GMT

执行cnvnator 报错，
ERROR in cling::CIFactory::createCI(): cannot extract standard library include paths!
Invoking:
x86_64-conda_cos6-linux-gnu-c++ -O3 -DNDEBUG -xc++ -E -v /dev/null 2>&1 | sed -n -e '/^.include/,${' -e '/^ /.*++/p' -e '}'
Results was:
With exit code 0
解决办法：
x86_64-conda_cos6-linux-gnu-c++ 这个程序在环境中
export PATH=$PATH:/opt/miniconda3/envs/cnvnator_env/bin/

svtools依赖python2.7 cnvnator 依赖python3.6 但是svtools会调用cnvnator

Reply to lumpy 结构突变分析流程 on Thu, 21 Oct 2021 06:04:22 GMT

anneng — Thu, 21 Oct 2021 06:04:22 GMT

CNVnator非常难安装用下面的conda环境
conda create -n cnvnator_env python=3.6
conda activate cnvnator_env
conda install -c conda-forge root_base=6.20
conda install -c conda-forge -c bioconda cnvnator

./cnvnator -root file.root -tree file.bam

nohup parallel "cnvnator -root root_libs/{= s:.[^.]+$::; s:.[^.]+$::; s:.[^.]+$::; s:.[^.]+$::; s:./::; =}.root -tree {}" ::: /ceph_disk2/yancaofenxi/raw_bam/ZGSP-.sv.alignment.sorted.bam &

Reply to lumpy 结构突变分析流程 on Wed, 20 Oct 2021 16:30:36 GMT

anneng — Wed, 20 Oct 2021 16:30:36 GMT

nohup parallel "svtyper -B {} -i all.gt.merged.with.f20.vcf > gt/{= s:.[^.]+$::; s:.[^.]+$::; s:.[^.]+$::; s:.[^.]+$::; s:./::; =}.vcf.tmp" ::: /ceph_disk2/yancaofenxi/raw_bam/ZGSP-.sv.alignment.sorted.bam &

通过.*/ 去掉路径

Reply to lumpy 结构突变分析流程 on Sat, 23 Oct 2021 17:31:35 GMT

anneng — Sat, 23 Oct 2021 17:31:35 GMT

验证svtools

cat gt.merged.vcf \
| vawk --header '{  $6="."; print }' \
| svtools genotype \
  -B ../raw_bam/ZGSP-1.sv.alignment.sorted.bam \
| sed 's/PR...=[0-9\.e,-]*\(;\)\{0,1\}\(\t\)\{0,1\}/\2/g' - \
> gt/ZGSP-1.vcf

parallel "svtyper -B {} -i gt.merged.vcf > gt/{= s:\.[^.]+$::; s:\.[^.]+$::; s:\.[^.]+$::; s:\.[^.]+$::; s:\..*/::; =}.vcf.tmp"  ::: ../raw_bam/ZGSP-*.sv.alignment.sorted.bam

需要将染色体独立成文件放到某个目录如chroms

#!/bin/bash
splitfa(){
 header=""
 while read line ; do
   if [ ${line:0:1} == ">" ] ; then
    header=$line
    echo "$header" >> ${header:1}.fa
   else
    seq="$line"
    echo "$seq" >> ${header:1}.fa
   fi
  done < $1
}

splitfa $1

nohup parallel "cnvnator -root {= s:\.[^.]+$::; s:\.[^.]+$::; s:\.[^.]+$::; s:\.[^.]+$::;  s:.*/::; =}.root -tree {}"  ::: /ceph_disk2/yancaofenxi/raw_bam/ZGSP-*.sv.alignment.sorted.bam &
nohup parallel "cnvnator -root {} -his 100 -d chroms"  ::: ../root_libs/ZGSP-*.root &
nohup parallel "cnvnator -root {} -his 1000 -d ../chroms"  ::: ZGSP-*.root &
nohup parallel "cnvnator -root {} -stat 100"  ::: ZGSP-*.root &
 nohup parallel "cnvnator -root {} -stat 1000"  ::: ZGSP-*.root &

nohup parallel "svtools copynumber --cnvnator /opt/miniconda3/envs/cnvnator_env/bin/cnvnator -s {=  s:\.[^.]+$::; s:\.[^.]+$::;  s:.*/::; =} -w 100 -r root_libs/{=  s:\.[^.]+$::; s:\.[^.]+$::;  s:.*/::; =}.root -c coordinates -i {} > cn/{=  s:\.[^.]+$::; s:\.[^.]+$::;  s:.*/::; =}.vcf"  ::: gt/*.vcf.tmp  &

注意：ROOT依赖下面的包

sudo apt-get install dpkg-dev cmake g++ gcc binutils libx11-dev libxpm-dev \
libxft-dev libxext-dev python libssl-dev

Reply to lumpy 结构突变分析流程 on Wed, 15 Sep 2021 07:02:47 GMT

anneng — Wed, 15 Sep 2021 07:02:47 GMT

svtyper比较慢用smoove试试

sudo docker run -d -v /ceph_disk3/sv_henan_yang/smoove/:/vcf -v /ceph_disk3/sv_henan_yang/raw_bam/:/bams brentp/smoove sh -c "ls /bams/ZGSP-*.sv.alignment.sorted.bam | xargs smoove genotype -p 10 --name prunus_sv --outdir /vcf --fasta /vcf/Prunus_persica_v2.0.a1_scaffolds.fasta --vcf /vcf/gt.merged.vcf "

Reply to lumpy 结构突变分析流程 on Tue, 07 Sep 2021 12:28:03 GMT

anneng — Tue, 07 Sep 2021 12:28:03 GMT

多个样本的处理方式
svtyper -B $(ls sample*.bam | paste -sd",") -i lumpy.raw.out.vcf > project.gt.vcf

Reply to lumpy 结构突变分析流程 on Fri, 03 Sep 2021 07:50:19 GMT

anneng — Fri, 03 Sep 2021 07:50:19 GMT

河南项目的过程

bwa mem -t 4 -M -R "@RG\tID:ZGSP-2\tPL:Illumina\tLB:ZGSP-2\tSM:ZGSP-2"  Prunus_persica_v2.0.a1_scaffolds.fasta ZGSP-2.R1.fq.gz ZGSP-2.R2.fq.gz  \
    | samblaster --excludeDups --addMateTags --maxSplitCount 2 --minNonOverlap 20 \
    | samtools view -S -b - \
    > sample.bam

samtools view -h ZGSP-683.alignment.sorted.markdup.bam      | /opt/lumpy-sv/scripts/extractSplitReads_BwaMem -i stdin     | samtools view -Sb -     > ZGSP-683.splitters.unsorted.bam
samtools sort ZGSP-683.split.unsorted.bam -o ZGSP-683.split.sorted.bam

samtools view -b -F 1294  ZGSP-683.alignment.sorted.markdup.bam > ZGSP-683.discordants.unsorted.bam

1462 nohup time lumpyexpress -B ../ZGSP-1.sv.alignment.sorted.bam -S ../ZGSP-1.split.sorted.bam -D ../ZGSP-1.discordants.sorted.bam -P -o 1.vcf >>1.log 2>&1 &
1468 nohup time lumpyexpress -B ../ZGSP-3.sv.alignment.sorted.bam -S ../ZGSP-3.split.sorted.bam -D ../ZGSP-3.discordants.sorted.bam -P -o 3.vcf >>2.log 2>&1 &
1493 samtools index ZGSP-1.sv.alignment.sorted.bam
1495 samtools index ZGSP-3.sv.alignment.sorted.bam
1510 nohup time svtyper -B ../ZGSP-1.sv.alignment.sorted.bam -S ../ZGSP-1.split.sorted.bam -i 1.vcf > 1.gt.svtyper.vcf 2>>sv.1.log &
1512 nohup time svtyper -B ../ZGSP-3.sv.alignment.sorted.bam -S ../ZGSP-3.split.sorted.bam -i 3.vcf > 3.gt.svtyper.vcf 2>>sv.3.log &
1528 svtools lsort 1.gt.svtyper.vcf 3.gt.svtyper.vcf >gt.sorted.vcf
1532 svtools lmerge -i gt.sorted.vcf >gt.merged.vcf
1535 svtyper -B ../ZGSP-1.sv.alignment.sorted.bam,-B ../ZGSP-3.sv.alignment.sorted.bam -i gt.merged.vcf >all.vcf
注意 svtyper多个样本要哦逗号分开不能是多个 -B 多个-B只去第一个样本