bwa-mem2和samtools:去宿主

bwa-mem2安装

https://github.com/bwa-mem2/bwa-mem2/releases

tar -xvjf bwa-mem2-2.2.1_x64-linux.tar.bz2
(完整路径)/bwa-mem2

 

 构建宿主index

bwa-mem2 index Host_genome_assembly.fasta(gz文件不行)

 

 

新增 5个文件

bwa-mem2比对去宿主

bwa-mem2 mem #看看各种参数
#看不太懂,看看教程用了什么参数
./bwa-mem2 mem -t 56 human_g1k_v37.fasta SRR7733443_1.fastq SRR7733443_2.fastq > d2_align.sam

非常的快呀 

Samtools安装

Releases · samtools/samtools · GitHub

tar -xvjf samtools-1.19.tar.bz2
./configure --prefix=(解压出的文件夹路径)
#把错误的东西补齐
make
make install
vim ~/.bashrc
#添加下述
export PATH=$PATH:绝对路径/samtools-1.19/bin
#保存退出
source ~/.bashrc
#完成
samtools

 使用samtools将宿主外的序列提取出来

#sam转bam
samtools view -Sb -@180 aligen.sam > aligen.bam
#sort
samtools sort -n -@180 aligen.bam -o aligen_sort.bam
#提取没比对到参考基因组的序列
samtools view -b -f 4 -@180 aligen_sort.bam > unmapped.bam
#转换为fastq
samtools bam2fq -1 unmapped_1.fastq -2 unmapped_2.fastq unmapped.bam
#bbtools的repair工具修复一下
bash repair.sh in=unmapped_1.fastq in2=unmapped_2.fastq out=unmapped_1_repaired.fastq out2=unmapped_2_repaired.fastq

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使用GATK的combinegvcf模块合并gvcf文件,可是到了这一步Using GATK jar /stor9000/apps/users/NWSUAF/2022050434/biosoft/gatk4.3/gatk-4.3.0.0/gatk-package-4.3.0.0-local.jar Running: java -Dsamjdk.use_async_io_read_samtools=false -Dsamjdk.use_async_io_write_samtools=true -Dsamjdk.use_async_io_write_tribble=false -Dsamjdk.compression_level=2 -jar /stor9000/apps/users/NWSUAF/2022050434/biosoft/gatk4.3/gatk-4.3.0.0/gatk-package-4.3.0.0-local.jar CombineGVCFs -R /stor9000/apps/users/NWSUAF/2008115251/genomes/ARS-UCD1.2_Btau5.0.1Y.fa --variant /stor9000/apps/users/NWSUAF/2020055419/home/xncattle/03.GVCF/01_out_GVCF/XN_22/1_XN_22.g.vcf.gz --variant /stor9000/apps/users/NWSUAF/2020055419/home/xncattle/03.GVCF/01_out_GVCF/XN_18/1_XN_18.g.vcf.gz -O /stor9000/apps/users/NWSUAF/2022050469/candy/bwa/gatk/Combine/chr1.g.vcf.gz 09:10:40.524 INFO NativeLibraryLoader - Loading libgkl_compression.so from jar:file:/stor9000/apps/users/NWSUAF/2022050434/biosoft/gatk4.3/gatk-4.3.0.0/gatk-package-4.3.0.0-local.jar!/com/intel/gkl/native/libgkl_compression.so 09:10:50.696 INFO CombineGVCFs - ------------------------------------------------------------ 09:10:50.697 INFO CombineGVCFs - The Genome Analysis Toolkit (GATK) v4.3.0.0 09:10:50.697 INFO CombineGVCFs - For support and documentation go to https://software.broadinstitute.org/gatk/ 09:10:50.698 INFO CombineGVCFs - Executing as 2022050469@node54 on Linux v3.10.0-1127.el7.x86_64 amd64 09:10:50.698 INFO CombineGVCFs - Java runtime: Java HotSpot(TM) 64-Bit Server VM v1.8.0_72-b15 09:10:50.698 INFO CombineGVCFs - Start Date/Time: July 21, 2023 9:10:40 AM CST 09:10:50.698 INFO CombineGVCFs - ------------------------------------------------------------ 09:10:50.698 INFO CombineGVCFs - ------------------------------------------------------------ 09:10:50.698 INFO CombineGVCFs - HTSJDK Version: 3.0.1 09:10:50.699 INFO CombineGVCFs - Picard Version: 2.27.5 09:10:50.699 INFO CombineGVCFs - Built for Spark Version: 2.4.5 09:10:50.699 INFO CombineGVCFs - HTSJDK Defaults.COMPRESSION_LEVEL : 2 09:10:50.699 INFO CombineGVCFs - HTSJDK Defa就停止了,没有输出文件,也没有报错文件
07-22
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