步骤如下:
1.上Genome Announcements网站(https://mra.asm.org/)找一篇细菌基因组文章;找到文章记载的SRA号
引用该篇文章的SRA号https://mra.asm.org/content/5/15/e00171-17
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2.从SRA数据库上用prefetch下载该文件
prefetch SRR5210995
得到如下结果
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3.Fastq-dump解压
fastq-dump --split-files ~/ncbi/public/sra/SRR5210995.sra
fastq-dump --gzip --split-files ~/ncbi/public/sra/SRR5210995.sra
结果如下
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4.对原始数据进行fastqc质量检测
fastqc SRR5210995_1.fastq.gz
fastqc SRR5210995_2.fastq.gz
结果如下
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去图形界面找到结果文件,打开
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5.原始数据过滤
mkdir trim_out
java -jar ~/Biosofts/Trimmomatic038/Trimmomatic-0.38/trimmomatic-0.38.jar PE -phred33 SRR5210995_1.fastq.gz SRR5210995_2.fastq.gz
./trim_out/output_forward_paired.fq.gz ./trirm_out/output_forward_unpaired.fq.gz ./trim_out/output_reverse_paired.fq.gz ./trim_out/output_reverse_unpaired.fq.gz ILLUMINACLIP:/home/eris/Biosofts/Trimmomatic038/Trimmomatic-0.38/adapters/TruSeq2-PE.fa:2:30:10 SLIDINGWINDOW:5:20 LEADING:20 TRAILING:20 MINLEN:75
结果如下
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6.再次查看质量
此时一定要选paired
cd /home/eris/trim_out
fastqc output_forward_paired.fq.gz
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再去图形界面查看
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7.运行SPAdes,组装细菌基因组
cd ~
spades.py --careful --pe1-1 SRR5210995_1.fastq.gz --pe1-2 SRR5210995_2.fastq.gz -o ./SPAdesout_SRR5210995
一定要确保自己的内存足够,不然会出现error:out of memory
结果图如下
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8.Quast评价组装的基因组效果
quast.py ~/SPAdesout_SRR5210995/contigs.fasta -o ~/SPAdesout_SRR5210995/quast_out
结果如下
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去图形界面下打开html
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