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修改脚本错误更新README.md

tags/v1.0
linzipeng 6 years ago
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README.md View File



### 甲基化原理简述 ### 甲基化原理简述


![](.\picture\BS原理.png)

全基因组亚硫酸氢盐测序用于研究基因粒度的DNA甲基化模式。 亚硫酸氢盐处理将胞嘧啶转化为尿嘧啶,但甲基化胞嘧啶不变。 全基因组亚硫酸氢盐测序用于研究基因粒度的DNA甲基化模式。 亚硫酸氢盐处理将胞嘧啶转化为尿嘧啶,但甲基化胞嘧啶不变。


比对软件(如Bismark)将基因组序列转化之后进行比对。 比对软件(如Bismark)将基因组序列转化之后进行比对。






![img](file:///C:/Users/monster/Documents/My Knowledge/temp/bbfd4716-e9eb-4735-9f7c-948838adbb62/128/index_files/004e3e03-4e75-487f-8be6-fa7509f07930.png)

### APP功能简述 ### APP功能简述


为了更好的分析全基因组甲基化数据,我们选用了目前最好的比对软件Bismark,构建了分析pipeline。用来提取全基因组的CpG,CHH,CHG甲基化模式信息。 为了更好的分析全基因组甲基化数据,我们选用了目前最好的比对软件Bismark,构建了分析pipeline。用来提取全基因组的CpG,CHH,CHG甲基化模式信息。


### WGBS分析流程 ### WGBS分析流程


![img](file:///C:/Users/monster/Documents/My Knowledge/temp/bbfd4716-e9eb-4735-9f7c-948838adbb62/128/index_files/c5901b21-4df4-46c4-a68a-11cb11476af7.png)


![](.\picture\WGBS流程.png)


### 参数说明 ### 参数说明




-p 多线程 -p 多线程


```





## 输入和输出 ## 输入和输出
以CpG_context_test_data_bismark_bt2.txt为例 以CpG_context_test_data_bismark_bt2.txt为例


``` ```

Bismark methylation extractor version v0.19.0 Bismark methylation extractor version v0.19.0
SRR15024317_length=86 - 1 57798691 z SRR15024317_length=86 - 1 57798691 z
SRR15024319_length=86 + 2 10166600 Z SRR15024319_length=86 + 2 10166600 Z
SRR15024331_length=86 + 11 77736289 Z SRR15024331_length=86 + 11 77736289 Z
SRR15024338_length=86 + 3 197272186 Z SRR15024338_length=86 + 3 197272186 Z

``` ```


第一行为Bismark的版本信息 第一行为Bismark的版本信息
不同字母表明不同的甲基化状态: 不同字母表明不同的甲基化状态:


``` ```

X 代表CHG中甲基化的C X 代表CHG中甲基化的C
x 代笔CHG中非甲基化的C x 代笔CHG中非甲基化的C
H 代表CHH中甲基化的C H 代表CHH中甲基化的C
z 代表CpG中非甲基化的C z 代表CpG中非甲基化的C
U 代表其他情况的甲基化C(CN或者CHN) U 代表其他情况的甲基化C(CN或者CHN)
u 代表其他情况的非甲基化C (CN或者CHN) u 代表其他情况的非甲基化C (CN或者CHN)

``` ```


#### 补充文件 #### 补充文件


上面的文件是methylation calling 最直接的证据,但是对于甲基化水平的定量来说,缺少了相关信息。运行bismark_methylation_extractor时,除了生成上述文件之外,还会有下列3个文件: 上面的文件是methylation calling 最直接的证据,但是对于甲基化水平的定量来说,缺少了相关信息。运行bismark_methylation_extractor时,除了生成上述文件之外,还会有下列3个文件:


```

test_data_bismark_bt2_splitting_report.txt test_data_bismark_bt2_splitting_report.txt
test_data_bismark_bt2.M-bias.txt test_data_bismark_bt2.M-bias.txt
test_data_bismark_bt2.M-bias_R1.png test_data_bismark_bt2.M-bias_R1.png




```



##### test_data_bismark_bt2_splitting_report.txt ##### test_data_bismark_bt2_splitting_report.txt


记录了该样本甲基化的汇总信息 记录了该样本甲基化的汇总信息


``` ```

Final Cytosine Methylation Report Final Cytosine Methylation Report
Total number of C’s analysed: 40348 Total number of C’s analysed: 40348
Total methylated C’s in CpG context: 1365 Total methylated C’s in CpG context: 1365
C methylated in CpG context: 66.8% C methylated in CpG context: 66.8%
C methylated in CHG context: 0.2% C methylated in CHG context: 0.2%
C methylated in CHH context: 0.4% C methylated in CHH context: 0.4%

``` ```


##### test_data_bismark_bt2.M-bias.txt ##### test_data_bismark_bt2.M-bias.txt
部分文件内容如下 部分文件内容如下


``` ```

CpG context CpG context
position count methylated count unmethylated % methylation coverage position count methylated count unmethylated % methylation coverage
1 42 13 76.36 55 1 42 13 76.36 55
2 31 9 77.50 40 2 31 9 77.50 40

``` ```





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inputs View File

{ {
"{{ project_name }}.fasta": "GRCh38.d1.vd1.fa",
"{{ project_name }}.ref_dir": "oss://pgx-reference-data/GRCh38.d1.vd1/",
"{{ project_name }}.ref_dir": "oss://pgx-test-data/wgbs/genome/hg38/",
"{{ project_name }}.fastq_1": "{{ read1 }}", "{{ project_name }}.fastq_1": "{{ read1 }}",
"{{ project_name }}.cluster_config": "{{ cluster if cluster != '' else 'OnDemand ecs.sn2ne.2xlarge img-ubuntu-vpc' }}", "{{ project_name }}.cluster_config": "{{ cluster if cluster != '' else 'OnDemand ecs.sn2ne.2xlarge img-ubuntu-vpc' }}",
"{{ project_name }}.docker": "registry.cn-shanghai.aliyuncs.com/pgx-docker-registry/sentieon-genomics:v2018.08.01",
"{{ project_name }}.docker": "registry-vpc.cn-shanghai.aliyuncs.com/pgx-docker-registry/bismark",
"{{ project_name }}.sample": "{{ sample_name }}", "{{ project_name }}.sample": "{{ sample_name }}",
"{{ project_name }}.disk_size": "{{ disk_size }}", "{{ project_name }}.disk_size": "{{ disk_size }}",
"{{ project_name }}.regions": "{{ regions }}",
"{{ project_name }}.fastq_2": "{{ read2 }}" "{{ project_name }}.fastq_2": "{{ read2 }}"
} }



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tasks/Dedup.wdl View File

command <<< command <<<
set -o pipefail set -o pipefail
set -e set -e
nt=$(nproc)
deduplicate_bismark -p --bam ${unsorted_bam} deduplicate_bismark -p --bam ${unsorted_bam}
>>> >>>
runtime { runtime {

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tasks/Dedup.wdl.bak View File

task Dedup {

String sample

File unsorted_bam

String docker
String cluster_config
String disk_size


command <<<
set -o pipefail
set -e
nt=$(nproc)
deduplicate_bismark -p --bam ${unsorted_bam}
>>>
runtime {
docker:docker
cluster: cluster_config
systemDisk: "cloud_ssd 40"
dataDisk: "cloud_ssd " + disk_size + " /cromwell_root/"
}

output {
File Dedup_bam = "${sample}_R1_val_1_bismark_bt2_pe.deduplicated.bam"
}
}







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tasks/Mapping.wdl View File

command <<< command <<<
set -o pipefail set -o pipefail
set -e set -e
nt=$(nproc)
bismark --bowtie2 -p ${nt} --bam ${ref_dir} -1 ${trim_read1} -2 ${trim_read2}
bismark --bowtie2 -p 8 --bam ${ref_dir} -1 ${trim_read1} -2 ${trim_read2}


>>> >>>



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tasks/Mapping.wdl.bak View File

task mapping {

File ref_dir
File trim_read1
File trim_read2

String sample
String docker
String cluster_config
String disk_size

command <<<
set -o pipefail
set -e
nt=$(nproc)
bismark --bowtie2 -p 8 --bam ${ref_dir} -1 ${trim_read1} -2 ${trim_read2}

>>>

runtime {
docker:docker
cluster: cluster_config
systemDisk: "cloud_ssd 40"
dataDisk: "cloud_ssd " + disk_size + " /cromwell_root/"
}
output {
File unsorted_bam = "${sample}_R1_val_1_bismark_bt2_pe.bam"
}
}

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