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day32 失败:EPIC-Seq流程

day32 失败:EPIC-Seq流程

作者: meraner | 来源:发表于2022-06-26 19:46 被阅读0次

    由于对cfDNA的兴趣,查到22年这篇在Nature biotechnology上的文章Inferring gene expression from cell-free DNA fragmentation profiles.想尝试跑一下EPIC-seq的流程。感谢文章作者提供的这么全面的资料。但,仍然很多不懂。

    无论用conda安装R,还是下载gz文件本地安装,都是失败。
    太难过了。
    一、 Rscript 这个是在Linux里面使用R要调用的命令。

    Rscript runEPIC.R –-bamdir  D:/work/research/ssDNA/result/ssDNA_1stsequence/bamdir –-tssinfo D:/work/research/ssDNA/result/ssDNA_1stsequence/code/priordata/all.tss.genes.canonical.ensembl75.txt  –-panelbed D:/work/research/ssDNA/result/ssDNA_1stsequence/code/priordata/all.tss.genes.canonical.ensembl75.txt.noheader –-targeted no  –-outdir D:/work/research/ssDNA/result/ssDNA_1stsequence/outdir  –-mapq 30 –-groupref <character> –-groupsize <double, number of genes per group> –-skipHist <binary, 0 or 1> –-skipOCFFrags <binary, 0 or 1>
    

    二、先建立虚拟环境
    conda create -n R_cfDNApipe python=3.8.3
    输入:conda activate R_cfDNApipe,报错。
    于是先输入:source activate,再输入 conda activate R_cfDNApipe

    image.png
    查看所有虚拟环境 conda env list
    image.png

    退出虚拟环境: conda deactivate
    参考教程: 虚拟环境
    虚拟环境:https://blog.csdn.net/robot8me/article/details/109471568
    conda的配置:https://www.bilibili.com/read/cv8956636
    conda瘦身:
    conda clean --packages --tarballs 删除缓存包,重新下载
    conda clean -i清除缓存,重新加载condo地址
    conda clean -p //删除没有用的包
    conda clean -t //删除tar包
    conda clean -y --all //删除所有的安装包及cache
    三、在Linux里直接安装R(直接安装失败,就是到PCRE那一步怎么都不成功)
    参考教程:
    https://www.136.la/jingpin/show-30899.html
    https://blog.csdn.net/tandelin/article/details/1183937531
    https://www.cnblogs.com/jessepeng/p/10984983.html
    下载文件 R-4.1.1.tar.gz,放到software目录下
    2,解压:tar -zxvf R-4.1.1.tar.gz
    3,进入解压后的文件夹
    进行configure,make, make install
    ./configure --prefix=/data/zds209/software/R --enable-R-shlib
    make
    make install
    4,configure遇到各种报错:

    1. zlib version >= 1.2.5需要安装
      由于公共software里有,只需直接.bashrc添加:
      export PATH=/data/software/zlib1.22.11/bin:$PATH
      export CFLAGS="-fPIC -I/data/software/zlib1.22.11/include"
      export LDFLAGS="-fPIC -L/data/software/zlib1.22.11/lib"

    2. bzip2需要安装
      下载https://sourceforge.net/projects/bzip2/files/latest/download

     tar xzvf bzip2-1.0.6.tar.gz
     cd bzip2-1.0.6
     make -f Makefile-libbz2_so
     make clean
     make
     make install PREFIX=/data/zds209/software/bzip2-1.0.6
    
    1. lzma version >= 5.0.3
      下载http://tukaani.org/xz/xz-5.2.3.tar.gz
    tar xzvf xz-5.2.3.tar.gz./configure --prefix=5. 
    cd tar xzvf xz-5.2.3.tar.gz
    ./configure --prefix=/data/zds209/software/xz-5.2.3
    make -j3
    make install
    
    1. PCRE >= 8.32
      下载 https://nchc.dl.sourceforge.net/project/pcre/pcre/8.42/pcre-8.42.tar.gz
    tar -zxvf pcre-8.42.tar.gz
    cd pcre-8.42
    
    ./configure --enable-utf8 --prefix=/data/zds209/software/pcre-8.42
    make -j3
    make install
    

    到.bashrc中

    export LC_ALL="zh_CN.UTF-8"
    export LANG="zh_CN.UTF-8"
    export PATH=/data/zds209/software/pcre-8.42/bin:$PATH
    export LD_LIBRARY_PATH=/data/software/pcre-8.42/bin:$LD_LIBRARY_PATH
    

    查看一下pcre的版本:
    zds209 07:14:44 ~pcre-config --version 8.44  查看UTF-8: zds209 07:15:09 ~locale
    LANG=zh_CN.UTF-8
    LC_CTYPE="zh_CN.UTF-8"
    LC_NUMERIC="zh_CN.UTF-8"
    LC_TIME="zh_CN.UTF-8"
    LC_COLLATE="zh_CN.UTF-8"
    LC_MONETARY="zh_CN.UTF-8"
    LC_MESSAGES="zh_CN.UTF-8"
    LC_PAPER="zh_CN.UTF-8"
    LC_NAME="zh_CN.UTF-8"
    LC_ADDRESS="zh_CN.UTF-8"
    LC_TELEPHONE="zh_CN.UTF-8"
    LC_MEASUREMENT="zh_CN.UTF-8"
    LC_IDENTIFICATION="zh_CN.UTF-8"
    LC_ALL=zh_CN.UTF-8

    用which查位置,发现不对劲,怎么conda里面也有。赶紧到.bashrc里面把conda的PATH注释掉。也不行。

    image.png
    于是进入到conda虚拟环境中,conda安装PCRE。

    到R-4.1.1目录下。

    ./configure --prefix=/data/zds209/software/R  --enable-R-shlib --with-readline=yes --with-libpng=yes --with-blas --with-pcre1 LDFLAGS="-L /data/zds209/software/bzip2-1.0.6/lib -L /data/zds209/software/xz-5.2.3/lib -L /data/zds209/software/bzip2-1.0.6/include -L /data/zds209/software/xz-5.2.3/include -L /data/zds209/software/pcre/lib -L /data/zds209/software/pcre/include" 
    
    四、虚拟环境中安装R
    https://www.jianshu.com/p/b9eb874fc8f4
    

    conda clean -i #清缓存
    conda create -n R_EPIC-seq python=3.8.3 #新建一个虚拟环境
    source activate
    conda activate R_EPIC-seq
    conda search R
    conda install r-base=4.0#总是在solving environment

    仍然失败。。。。。
    
    
    
    教程:
    https://yunwei365.blog.csdn.net/article/details/108126550?spm=1001.2101.3001.6650.12&utm_medium=distribute.pc_relevant.none-task-blog-2~default~OPENSEARCH~default-12.no_search_link&depth_1-utm_source=distribute.pc_relevant.none-task-blog-2~default~OPENSEARCH~default-12.no_search_link&utm_relevant_index=18
    
    
    
    
    
    
    
    
    

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