運行環境:
conda 4.10.3
conda create -n chipseq python=3.6
conda install -c bioconda parallel-fastq-dump
conda install -y trim-galore
數據文件:從NCBI上下載
wget https://sra-pub-run-odp.s3.amazonaws.com/sra/SRR8557351/SRR8557351
將該數據下載后,經過parallel-fastq-dump轉化成fastq.gz
mv SRR8557351 IP_H33K
mkdir fastq
parallel-fastq-dump --sra-id IP_H33K --threads 10 --outdir fastq/ --split-files --gzip
進行fastqc質控
mkdir fastqc
fastqc -o fastqc/ -t 10 IP_H33K_1.fastq.gz
出現了如下報錯:
/home/miniconda3/envs/chipseq/bin/java: 8: Syntax error: Unterminated quoted string
咨詢大佬后表示可能是java編譯環境出問題,軟件與java環境不兼容
卸載java和miniconda后重新安裝最新版本java和anaconda,報錯依舊存在
仔細查看下parallel-fastq-dump和trim-galore軟件中的java相應依賴包的文件,發現版本原來是之前清華鏡像源上的,盡管能下載,但是下載下來的版本沖突,更換channels
conda config --add channels conda-forge
conda config --add channels defaults
conda config --add channels r
conda config --add channels bioconda
conda config --add channels https://mirrors.tuna.tsinghua.edu.cn/anaconda/pkgs/free/
更換后重新走一遍上面的流程,盡管fastqc可以運行,但是同樣報錯:undefined symbol:FT_Done_MM_Var
這個問題就好解決了,應該是libfontconfig.so.1版本的問題,具體解決方式簡書上可以輕松搜索到,
更換軟連接為libfontconfig.so.1.9后解決。
再次嘗試fastqc
Started analysis of IP_H33K_1.fastq.gz
Approx 5% complete for IP_H33K_1.fastq.gz
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Analysis complete for IP_H33K_1.fastq.gz
(chipseq) root@ubuntu:/test/fastq#
搞定!
(補充:這類問題后來詢問了個做linux的大佬,大佬的回應是虛擬機太菜了,因為本人是之前用的閹割版虛擬機,后來裝了VMware,用的ubuntu18.4版本,anaconda也安裝了一個比較穩定的4.8.4版本,運行起來更為流暢,不得不說閹割版虛擬機害人不淺呀!)