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### 甲基化原理简述 |
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### 甲基化原理简述 |
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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。 |
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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。 |
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比对软件(如Bismark)将基因组序列转化之后进行比对。 |
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比对软件(如Bismark)将基因组序列转化之后进行比对。 |
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### APP功能简述 |
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### APP功能简述 |
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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。 |
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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。 |
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### WGBS分析流程 |
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### WGBS分析流程 |
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### 参数说明 |
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### 参数说明 |
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-p 多线程 |
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-p 多线程 |
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``` |
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## 输入和输出 |
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## 输入和输出 |
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以CpG_context_test_data_bismark_bt2.txt为例 |
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以CpG_context_test_data_bismark_bt2.txt为例 |
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``` |
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``` |
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Bismark methylation extractor version v0.19.0 |
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Bismark methylation extractor version v0.19.0 |
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SRR15024317_length=86 - 1 57798691 z |
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SRR15024317_length=86 - 1 57798691 z |
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SRR15024319_length=86 + 2 10166600 Z |
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SRR15024319_length=86 + 2 10166600 Z |
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SRR15024331_length=86 + 11 77736289 Z |
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SRR15024331_length=86 + 11 77736289 Z |
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SRR15024338_length=86 + 3 197272186 Z |
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SRR15024338_length=86 + 3 197272186 Z |
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``` |
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``` |
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第一行为Bismark的版本信息 |
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第一行为Bismark的版本信息 |
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不同字母表明不同的甲基化状态: |
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不同字母表明不同的甲基化状态: |
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``` |
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``` |
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X 代表CHG中甲基化的C |
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X 代表CHG中甲基化的C |
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x 代笔CHG中非甲基化的C |
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x 代笔CHG中非甲基化的C |
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H 代表CHH中甲基化的C |
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H 代表CHH中甲基化的C |
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z 代表CpG中非甲基化的C |
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z 代表CpG中非甲基化的C |
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U 代表其他情况的甲基化C(CN或者CHN) |
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U 代表其他情况的甲基化C(CN或者CHN) |
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u 代表其他情况的非甲基化C (CN或者CHN) |
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u 代表其他情况的非甲基化C (CN或者CHN) |
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``` |
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``` |
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#### 补充文件 |
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#### 补充文件 |
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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: |
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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: |
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``` |
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test_data_bismark_bt2_splitting_report.txt |
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test_data_bismark_bt2_splitting_report.txt |
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test_data_bismark_bt2.M-bias.txt |
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test_data_bismark_bt2.M-bias.txt |
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test_data_bismark_bt2.M-bias_R1.png |
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test_data_bismark_bt2.M-bias_R1.png |
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``` |
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##### test_data_bismark_bt2_splitting_report.txt |
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##### test_data_bismark_bt2_splitting_report.txt |
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记录了该样本甲基化的汇总信息 |
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记录了该样本甲基化的汇总信息 |
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``` |
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``` |
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Final Cytosine Methylation Report |
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Final Cytosine Methylation Report |
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Total number of C’s analysed: 40348 |
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Total number of C’s analysed: 40348 |
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Total methylated C’s in CpG context: 1365 |
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Total methylated C’s in CpG context: 1365 |
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C methylated in CpG context: 66.8% |
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C methylated in CpG context: 66.8% |
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C methylated in CHG context: 0.2% |
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C methylated in CHG context: 0.2% |
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C methylated in CHH context: 0.4% |
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C methylated in CHH context: 0.4% |
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``` |
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``` |
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##### test_data_bismark_bt2.M-bias.txt |
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##### test_data_bismark_bt2.M-bias.txt |
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部分文件内容如下 |
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部分文件内容如下 |
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``` |
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``` |
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CpG context |
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CpG context |
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position count methylated count unmethylated % methylation coverage |
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position count methylated count unmethylated % methylation coverage |
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1 42 13 76.36 55 |
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1 42 13 76.36 55 |
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2 31 9 77.50 40 |
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2 31 9 77.50 40 |
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``` |
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``` |
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