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  133. <h1 id="seo-header">inferCNV推断肿瘤染色体变异</h1>
  134. <div class="markdown-body">
  135. <h2 id="第一步-获取对应物种的基因位置注释文件"><a href="#第一步-获取对应物种的基因位置注释文件" class="headerlink" title="第一步 获取对应物种的基因位置注释文件"></a>第一步 获取对应物种的基因位置注释文件</h2><ul>
  136. <li><a target="_blank" rel="noopener" href="https://www.gencodegenes.org/human/">https://www.gencodegenes.org/human/</a></li>
  137. <li>wget <a target="_blank" rel="noopener" href="https://ftp.ebi.ac.uk/pub/databases/gencode/Gencode/_human/release/_39/gencode.v39.chr/_patch/_hapl/_scaff.annotation.gtf.gz">https://ftp.ebi.ac.uk/pub/databases/gencode/Gencode\_human/release\_39/gencode.v39.chr\_patch\_hapl\_scaff.annotation.gtf.gz</a></li>
  138. <li>gunzip gencode.v39.chr_patch_hapl_scaff.annotation.gtf.gz</li>
  139. <li>hg39 &lt;- read.table(‘gencode.v39.chr_patch_hapl_scaff.annotation.gtf’, sep &#x3D; ‘\t’, comment.char &#x3D; “#”)</li>
  140. <li>hg39[‘gene_id’] &lt;- stringr::str_extract(hg39$V9,”(?&lt;&#x3D;gene_id ).+?(?&#x3D;;)”)</li>
  141. <li>hg39[‘gene_name’] &lt;- stringr::str_extract(hg39$V9,”(?&lt;&#x3D;gene_name ).+?(?&#x3D;;)”)</li>
  142. <li>或者直接使用TCGA的基因位置注释:<a target="_blank" rel="noopener" href="https://file-cdn.limour.top/bix/22.02.18.tcga_rowRanges.csv.gz">https://file-cdn.limour.top/bix/22.02.18.tcga_rowRanges.csv.gz</a></li>
  143. </ul>
  144. <h2 id="第二步-获取分析数据的基因位置信息"><a href="#第二步-获取分析数据的基因位置信息" class="headerlink" title="第二步 获取分析数据的基因位置信息"></a>第二步 获取分析数据的基因位置信息</h2><figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br><span class="line">6</span><br><span class="line">7</span><br><span class="line">8</span><br><span class="line">9</span><br><span class="line">10</span><br></pre></td><td class="code"><pre><code class="hljs r">f_df_order <span class="hljs-operator">&lt;-</span> <span class="hljs-keyword">function</span><span class="hljs-punctuation">(</span>df<span class="hljs-punctuation">,</span> oN<span class="hljs-punctuation">,</span> groupN<span class="hljs-punctuation">,</span> decreasing <span class="hljs-operator">=</span> <span class="hljs-built_in">T</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">&#123;</span><br> df <span class="hljs-operator">&lt;-</span> as.data.frame<span class="hljs-punctuation">(</span>df<span class="hljs-punctuation">)</span><br> <span class="hljs-keyword">for</span><span class="hljs-punctuation">(</span>oi <span class="hljs-keyword">in</span> levels<span class="hljs-punctuation">(</span>df<span class="hljs-punctuation">[[</span>groupN<span class="hljs-punctuation">]</span><span class="hljs-punctuation">]</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">&#123;</span><br> idx <span class="hljs-operator">&lt;-</span> df<span class="hljs-punctuation">[[</span>groupN<span class="hljs-punctuation">]</span><span class="hljs-punctuation">]</span> <span class="hljs-operator">==</span> oi<br> tmp <span class="hljs-operator">&lt;-</span> df<span class="hljs-punctuation">[</span>idx<span class="hljs-punctuation">,</span> <span class="hljs-punctuation">]</span><br> tmp <span class="hljs-operator">&lt;-</span> tmp<span class="hljs-punctuation">[</span>order<span class="hljs-punctuation">(</span>tmp<span class="hljs-punctuation">[[</span>oN<span class="hljs-punctuation">]</span><span class="hljs-punctuation">]</span><span class="hljs-punctuation">,</span> decreasing <span class="hljs-operator">=</span> decreasing<span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><span class="hljs-punctuation">]</span><br> df<span class="hljs-punctuation">[</span>idx<span class="hljs-punctuation">,</span> <span class="hljs-punctuation">]</span> <span class="hljs-operator">&lt;-</span> tmp<br> <span class="hljs-punctuation">&#125;</span><br> df<br><span class="hljs-punctuation">&#125;</span><br></code></pre></td></tr></table></figure>
  145. <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br></pre></td><td class="code"><pre><code class="hljs r">tcga_gene <span class="hljs-operator">&lt;-</span> read.csv<span class="hljs-punctuation">(</span><span class="hljs-string">&#x27;22.02.18.tcga_rowRanges.csv&#x27;</span><span class="hljs-punctuation">,</span> row.names <span class="hljs-operator">=</span> <span class="hljs-number">1</span><span class="hljs-punctuation">)</span><br>tcga_gene<span class="hljs-punctuation">[</span><span class="hljs-string">&#x27;seqnames&#x27;</span><span class="hljs-punctuation">]</span> <span class="hljs-operator">&lt;-</span> factor<span class="hljs-punctuation">(</span>tcga_gene<span class="hljs-operator">$</span>seqnames<span class="hljs-punctuation">,</span> levels <span class="hljs-operator">=</span> paste0<span class="hljs-punctuation">(</span><span class="hljs-string">&#x27;chr&#x27;</span><span class="hljs-punctuation">,</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-number">1</span><span class="hljs-operator">:</span><span class="hljs-number">22</span><span class="hljs-punctuation">,</span><span class="hljs-string">&#x27;X&#x27;</span><span class="hljs-punctuation">,</span> <span class="hljs-string">&#x27;Y&#x27;</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br>tcga_gene <span class="hljs-operator">&lt;-</span> tcga_gene<span class="hljs-punctuation">[</span>order<span class="hljs-punctuation">(</span><span class="hljs-built_in">as.numeric</span><span class="hljs-punctuation">(</span>tcga_gene<span class="hljs-operator">$</span>seqnames<span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><span class="hljs-punctuation">]</span><br>tcga_gene <span class="hljs-operator">&lt;-</span> f_df_order<span class="hljs-punctuation">(</span>tcga_gene<span class="hljs-punctuation">,</span> <span class="hljs-string">&#x27;start&#x27;</span><span class="hljs-punctuation">,</span> <span class="hljs-string">&#x27;seqnames&#x27;</span><span class="hljs-punctuation">,</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">)</span><br></code></pre></td></tr></table></figure>
  146. <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br></pre></td><td class="code"><pre><code class="hljs r">sce <span class="hljs-operator">&lt;-</span> readRDS<span class="hljs-punctuation">(</span><span class="hljs-string">&#x27;~/upload/zl_liu/data/pca.rds&#x27;</span><span class="hljs-punctuation">)</span><br>sce<span class="hljs-operator">@</span>meta.data <span class="hljs-operator">&lt;-</span> readRDS<span class="hljs-punctuation">(</span><span class="hljs-string">&#x27;../fig1/E_meta.data.rds&#x27;</span><span class="hljs-punctuation">)</span><br>tcga_gene <span class="hljs-operator">&lt;-</span> tcga_gene<span class="hljs-punctuation">[</span>tcga_gene<span class="hljs-operator">$</span>external_gene_name <span class="hljs-operator">%in%</span> rownames<span class="hljs-punctuation">(</span>sce<span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">&#x27;external_gene_name&#x27;</span><span class="hljs-punctuation">,</span> <span class="hljs-string">&#x27;seqnames&#x27;</span><span class="hljs-punctuation">,</span> <span class="hljs-string">&#x27;start&#x27;</span><span class="hljs-punctuation">,</span> <span class="hljs-string">&#x27;end&#x27;</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">]</span><br>write.table<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> tcga_gene<span class="hljs-punctuation">,</span> file <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;gencode_downsampled.txt&#x27;</span><span class="hljs-punctuation">,</span> sep <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;\t&#x27;</span><span class="hljs-punctuation">,</span> row.names <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">,</span> col.names <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">,</span> <span class="hljs-built_in">quote</span> <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">)</span><br></code></pre></td></tr></table></figure>
  147. <h2 id="第三步-导出-raw-counts-matrix-和-annotations-file"><a href="#第三步-导出-raw-counts-matrix-和-annotations-file" class="headerlink" title="第三步 导出 raw_counts_matrix 和 annotations_file"></a>第三步 导出 raw_counts_matrix 和 annotations_file</h2><figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br></pre></td><td class="code"><pre><code class="hljs r">library<span class="hljs-punctuation">(</span>Seurat<span class="hljs-punctuation">)</span><br>sce <span class="hljs-operator">&lt;-</span> readRDS<span class="hljs-punctuation">(</span><span class="hljs-string">&#x27;~/upload/zl_liu/data/pca.rds&#x27;</span><span class="hljs-punctuation">)</span><br>sce<span class="hljs-operator">@</span>meta.data <span class="hljs-operator">&lt;-</span> readRDS<span class="hljs-punctuation">(</span><span class="hljs-string">&#x27;../fig1/E_meta.data.rds&#x27;</span><span class="hljs-punctuation">)</span><br>counts_matrix <span class="hljs-operator">=</span> GetAssayData<span class="hljs-punctuation">(</span>seurat_obj<span class="hljs-punctuation">,</span> slot<span class="hljs-operator">=</span><span class="hljs-string">&quot;counts&quot;</span><span class="hljs-punctuation">)</span><br>saveRDS<span class="hljs-punctuation">(</span><span class="hljs-built_in">round</span><span class="hljs-punctuation">(</span>counts_matrix<span class="hljs-punctuation">,</span> digits<span class="hljs-operator">=</span><span class="hljs-number">3</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> <span class="hljs-string">&quot;sc.10x.counts.matrix&quot;</span><span class="hljs-punctuation">)</span><br></code></pre></td></tr></table></figure>
  148. <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br><span class="line">6</span><br><span class="line">7</span><br><span class="line">8</span><br><span class="line">9</span><br><span class="line">10</span><br><span class="line">11</span><br><span class="line">12</span><br><span class="line">13</span><br><span class="line">14</span><br></pre></td><td class="code"><pre><code class="hljs r">df <span class="hljs-operator">&lt;-</span> sce<span class="hljs-operator">@</span>meta.data<br>df <span class="hljs-operator">&lt;-</span> subset<span class="hljs-punctuation">(</span>df<span class="hljs-punctuation">,</span> cell_type_fig1A <span class="hljs-operator">%in%</span> <span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">&#x27;Fibroblasts&#x27;</span><span class="hljs-punctuation">,</span> <span class="hljs-string">&#x27;Luminal&#x27;</span><span class="hljs-punctuation">,</span> <span class="hljs-string">&#x27;Endothelial&#x27;</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">)</span><br><br>tmp <span class="hljs-operator">&lt;-</span> subset<span class="hljs-punctuation">(</span>df<span class="hljs-punctuation">,</span> patient_id <span class="hljs-operator">==</span> <span class="hljs-string">&#x27;patient1&#x27;</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">[</span><span class="hljs-string">&#x27;cell_type_fig1A&#x27;</span><span class="hljs-punctuation">]</span><br>write.table<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> tmp<span class="hljs-punctuation">,</span> file <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;annotations_file_p1.txt&#x27;</span><span class="hljs-punctuation">,</span> sep <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;\t&#x27;</span><span class="hljs-punctuation">,</span> row.names <span class="hljs-operator">=</span> <span class="hljs-built_in">T</span><span class="hljs-punctuation">,</span> col.names <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">,</span> <span class="hljs-built_in">quote</span> <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">)</span><br><br>tmp <span class="hljs-operator">&lt;-</span> subset<span class="hljs-punctuation">(</span>df<span class="hljs-punctuation">,</span> patient_id <span class="hljs-operator">==</span> <span class="hljs-string">&#x27;patient3&#x27;</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">[</span><span class="hljs-string">&#x27;cell_type_fig1A&#x27;</span><span class="hljs-punctuation">]</span><br>write.table<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> tmp<span class="hljs-punctuation">,</span> file <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;annotations_file_p3.txt&#x27;</span><span class="hljs-punctuation">,</span> sep <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;\t&#x27;</span><span class="hljs-punctuation">,</span> row.names <span class="hljs-operator">=</span> <span class="hljs-built_in">T</span><span class="hljs-punctuation">,</span> col.names <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">,</span> <span class="hljs-built_in">quote</span> <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">)</span><br><br>tmp <span class="hljs-operator">&lt;-</span> subset<span class="hljs-punctuation">(</span>df<span class="hljs-punctuation">,</span> patient_id <span class="hljs-operator">==</span> <span class="hljs-string">&#x27;patient4&#x27;</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">[</span><span class="hljs-string">&#x27;cell_type_fig1A&#x27;</span><span class="hljs-punctuation">]</span><br>write.table<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> tmp<span class="hljs-punctuation">,</span> file <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;annotations_file_p4.txt&#x27;</span><span class="hljs-punctuation">,</span> sep <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;\t&#x27;</span><span class="hljs-punctuation">,</span> row.names <span class="hljs-operator">=</span> <span class="hljs-built_in">T</span><span class="hljs-punctuation">,</span> col.names <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">,</span> <span class="hljs-built_in">quote</span> <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">)</span><br><br>tmp <span class="hljs-operator">&lt;-</span> subset<span class="hljs-punctuation">(</span>df<span class="hljs-punctuation">,</span> patient_id <span class="hljs-operator">==</span> <span class="hljs-string">&#x27;patient5&#x27;</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">[</span><span class="hljs-string">&#x27;cell_type_fig1A&#x27;</span><span class="hljs-punctuation">]</span><br>write.table<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> tmp<span class="hljs-punctuation">,</span> file <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;annotations_file_p5.txt&#x27;</span><span class="hljs-punctuation">,</span> sep <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;\t&#x27;</span><span class="hljs-punctuation">,</span> row.names <span class="hljs-operator">=</span> <span class="hljs-built_in">T</span><span class="hljs-punctuation">,</span> col.names <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">,</span> <span class="hljs-built_in">quote</span> <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">)</span><br></code></pre></td></tr></table></figure>
  149. <h2 id="第四步-进行分析"><a href="#第四步-进行分析" class="headerlink" title="第四步 进行分析"></a>第四步 进行分析</h2><figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br></pre></td><td class="code"><pre><code class="hljs r">fuck <span class="hljs-operator">&lt;-</span> read.table<span class="hljs-punctuation">(</span><span class="hljs-string">&#x27;gencode_downsampled.txt&#x27;</span><span class="hljs-punctuation">,</span> sep <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;\t&#x27;</span><span class="hljs-punctuation">)</span><br>fuck <span class="hljs-operator">&lt;-</span> fuck<span class="hljs-punctuation">[</span><span class="hljs-operator">!</span>duplicated<span class="hljs-punctuation">(</span>fuck<span class="hljs-operator">$</span>V1<span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><span class="hljs-punctuation">]</span><br>write.table<span class="hljs-punctuation">(</span>x <span class="hljs-operator">=</span> fuck<span class="hljs-punctuation">,</span> file <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;gencode_downsampled.txt&#x27;</span><span class="hljs-punctuation">,</span> sep <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;\t&#x27;</span><span class="hljs-punctuation">,</span> row.names <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">,</span> col.names <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">,</span> <span class="hljs-built_in">quote</span> <span class="hljs-operator">=</span> <span class="hljs-built_in">F</span><span class="hljs-punctuation">)</span><br></code></pre></td></tr></table></figure>
  150. <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br><span class="line">6</span><br><span class="line">7</span><br><span class="line">8</span><br><span class="line">9</span><br><span class="line">10</span><br><span class="line">11</span><br><span class="line">12</span><br><span class="line">13</span><br><span class="line">14</span><br><span class="line">15</span><br><span class="line">16</span><br><span class="line">17</span><br><span class="line">18</span><br><span class="line">19</span><br><span class="line">20</span><br><span class="line">21</span><br><span class="line">22</span><br><span class="line">23</span><br><span class="line">24</span><br></pre></td><td class="code"><pre><code class="hljs r">infercnv_obj_p1 <span class="hljs-operator">=</span> CreateInfercnvObject<span class="hljs-punctuation">(</span>raw_counts_matrix<span class="hljs-operator">=</span>readRDS<span class="hljs-punctuation">(</span><span class="hljs-string">&quot;sc.10x.counts.matrix&quot;</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> annotations_file<span class="hljs-operator">=</span><span class="hljs-string">&quot;annotations_file_p1.txt&quot;</span><span class="hljs-punctuation">,</span><br> delim<span class="hljs-operator">=</span><span class="hljs-string">&quot;\t&quot;</span><span class="hljs-punctuation">,</span><br> gene_order_file<span class="hljs-operator">=</span><span class="hljs-string">&quot;gencode_downsampled.txt&quot;</span><span class="hljs-punctuation">,</span><br> ref_group_names<span class="hljs-operator">=</span><span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">&#x27;Fibroblasts&#x27;</span><span class="hljs-punctuation">,</span> <span class="hljs-string">&#x27;Endothelial&#x27;</span><span class="hljs-punctuation">)</span> <br> <span class="hljs-punctuation">)</span><br>infercnv_obj_p3 <span class="hljs-operator">=</span> CreateInfercnvObject<span class="hljs-punctuation">(</span>raw_counts_matrix<span class="hljs-operator">=</span>readRDS<span class="hljs-punctuation">(</span><span class="hljs-string">&quot;sc.10x.counts.matrix&quot;</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> annotations_file<span class="hljs-operator">=</span><span class="hljs-string">&quot;annotations_file_p3.txt&quot;</span><span class="hljs-punctuation">,</span><br> delim<span class="hljs-operator">=</span><span class="hljs-string">&quot;\t&quot;</span><span class="hljs-punctuation">,</span><br> gene_order_file<span class="hljs-operator">=</span><span class="hljs-string">&quot;gencode_downsampled.txt&quot;</span><span class="hljs-punctuation">,</span><br> ref_group_names<span class="hljs-operator">=</span><span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">&#x27;Fibroblasts&#x27;</span><span class="hljs-punctuation">,</span> <span class="hljs-string">&#x27;Endothelial&#x27;</span><span class="hljs-punctuation">)</span> <br> <span class="hljs-punctuation">)</span><br>infercnv_obj_p4 <span class="hljs-operator">=</span> CreateInfercnvObject<span class="hljs-punctuation">(</span>raw_counts_matrix<span class="hljs-operator">=</span>readRDS<span class="hljs-punctuation">(</span><span class="hljs-string">&quot;sc.10x.counts.matrix&quot;</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> annotations_file<span class="hljs-operator">=</span><span class="hljs-string">&quot;annotations_file_p4.txt&quot;</span><span class="hljs-punctuation">,</span><br> delim<span class="hljs-operator">=</span><span class="hljs-string">&quot;\t&quot;</span><span class="hljs-punctuation">,</span><br> gene_order_file<span class="hljs-operator">=</span><span class="hljs-string">&quot;gencode_downsampled.txt&quot;</span><span class="hljs-punctuation">,</span><br> ref_group_names<span class="hljs-operator">=</span><span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">&#x27;Fibroblasts&#x27;</span><span class="hljs-punctuation">,</span> <span class="hljs-string">&#x27;Endothelial&#x27;</span><span class="hljs-punctuation">)</span> <br> <span class="hljs-punctuation">)</span><br>infercnv_obj_p5 <span class="hljs-operator">=</span> CreateInfercnvObject<span class="hljs-punctuation">(</span>raw_counts_matrix<span class="hljs-operator">=</span>readRDS<span class="hljs-punctuation">(</span><span class="hljs-string">&quot;sc.10x.counts.matrix&quot;</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span><br> annotations_file<span class="hljs-operator">=</span><span class="hljs-string">&quot;annotations_file_p5.txt&quot;</span><span class="hljs-punctuation">,</span><br> delim<span class="hljs-operator">=</span><span class="hljs-string">&quot;\t&quot;</span><span class="hljs-punctuation">,</span><br> gene_order_file<span class="hljs-operator">=</span><span class="hljs-string">&quot;gencode_downsampled.txt&quot;</span><span class="hljs-punctuation">,</span><br> ref_group_names<span class="hljs-operator">=</span><span class="hljs-built_in">c</span><span class="hljs-punctuation">(</span><span class="hljs-string">&#x27;Fibroblasts&#x27;</span><span class="hljs-punctuation">,</span> <span class="hljs-string">&#x27;Endothelial&#x27;</span><span class="hljs-punctuation">)</span> <br> <span class="hljs-punctuation">)</span><br></code></pre></td></tr></table></figure>
  151. <figure class="highlight r"><table><tr><td class="gutter"><pre><span class="line">1</span><br><span class="line">2</span><br><span class="line">3</span><br><span class="line">4</span><br><span class="line">5</span><br><span class="line">6</span><br><span class="line">7</span><br><span class="line">8</span><br><span class="line">9</span><br><span class="line">10</span><br><span class="line">11</span><br><span class="line">12</span><br><span class="line">13</span><br><span class="line">14</span><br><span class="line">15</span><br><span class="line">16</span><br><span class="line">17</span><br><span class="line">18</span><br><span class="line">19</span><br><span class="line">20</span><br><span class="line">21</span><br><span class="line">22</span><br><span class="line">23</span><br><span class="line">24</span><br><span class="line">25</span><br><span class="line">26</span><br><span class="line">27</span><br><span class="line">28</span><br><span class="line">29</span><br><span class="line">30</span><br><span class="line">31</span><br><span class="line">32</span><br><span class="line">33</span><br><span class="line">34</span><br><span class="line">35</span><br><span class="line">36</span><br><span class="line">37</span><br><span class="line">38</span><br><span class="line">39</span><br><span class="line">40</span><br><span class="line">41</span><br><span class="line">42</span><br><span class="line">43</span><br><span class="line">44</span><br><span class="line">45</span><br><span class="line">46</span><br><span class="line">47</span><br></pre></td><td class="code"><pre><code class="hljs r">infercnv_obj_p1 <span class="hljs-operator">=</span> infercnv<span class="hljs-operator">::</span>run<span class="hljs-punctuation">(</span>infercnv_obj_p1<span class="hljs-punctuation">,</span><br> cutoff<span class="hljs-operator">=</span><span class="hljs-number">0.1</span><span class="hljs-punctuation">,</span> <span class="hljs-comment"># use 1 for smart-seq, 0.1 for 10x-genomics</span><br> out_dir<span class="hljs-operator">=</span><span class="hljs-string">&quot;p1&quot;</span><span class="hljs-punctuation">,</span> <span class="hljs-comment"># dir is auto-created for storing outputs</span><br> cluster_by_groups<span class="hljs-operator">=</span><span class="hljs-built_in">F</span><span class="hljs-punctuation">,</span> <br> analysis_mode<span class="hljs-operator">=</span><span class="hljs-string">&quot;subclusters&quot;</span><span class="hljs-punctuation">,</span><br> tumor_subcluster_partition_method <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;random_trees&#x27;</span><span class="hljs-punctuation">,</span><br> denoise<span class="hljs-operator">=</span><span class="hljs-literal">TRUE</span><span class="hljs-punctuation">,</span><br> HMM<span class="hljs-operator">=</span><span class="hljs-literal">TRUE</span><span class="hljs-punctuation">,</span><br> num_threads<span class="hljs-operator">=</span><span class="hljs-number">12</span><span class="hljs-punctuation">)</span><br>saveRDS<span class="hljs-punctuation">(</span>infercnv_obj_p1<span class="hljs-punctuation">,</span> <span class="hljs-string">&#x27;infercnv_obj_p1.rds&#x27;</span><span class="hljs-punctuation">)</span><br>system<span class="hljs-punctuation">(</span>paste<span class="hljs-punctuation">(</span><span class="hljs-string">&quot;/opt/conda/bin/python3 /home/jovyan/upload/zl_liu/wecomchan.py&quot;</span><span class="hljs-punctuation">,</span> <span class="hljs-string">&quot;&#x27;task p1 successfully completed&#x27;&quot;</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> intern <span class="hljs-operator">=</span> <span class="hljs-built_in">T</span><span class="hljs-punctuation">)</span><br><br>infercnv_obj_p3 <span class="hljs-operator">=</span> infercnv<span class="hljs-operator">::</span>run<span class="hljs-punctuation">(</span>infercnv_obj_p3<span class="hljs-punctuation">,</span><br> cutoff<span class="hljs-operator">=</span><span class="hljs-number">0.1</span><span class="hljs-punctuation">,</span> <span class="hljs-comment"># use 1 for smart-seq, 0.1 for 10x-genomics</span><br> out_dir<span class="hljs-operator">=</span><span class="hljs-string">&quot;p3&quot;</span><span class="hljs-punctuation">,</span> <span class="hljs-comment"># dir is auto-created for storing outputs</span><br> cluster_by_groups<span class="hljs-operator">=</span><span class="hljs-built_in">F</span><span class="hljs-punctuation">,</span> <br> analysis_mode<span class="hljs-operator">=</span><span class="hljs-string">&quot;subclusters&quot;</span><span class="hljs-punctuation">,</span><br> tumor_subcluster_partition_method <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;random_trees&#x27;</span><span class="hljs-punctuation">,</span><br> denoise<span class="hljs-operator">=</span><span class="hljs-literal">TRUE</span><span class="hljs-punctuation">,</span><br> HMM<span class="hljs-operator">=</span><span class="hljs-literal">TRUE</span><span class="hljs-punctuation">,</span><br> num_threads<span class="hljs-operator">=</span><span class="hljs-number">12</span><span class="hljs-punctuation">)</span><br>saveRDS<span class="hljs-punctuation">(</span>infercnv_obj_p3<span class="hljs-punctuation">,</span> <span class="hljs-string">&#x27;infercnv_obj_p3.rds&#x27;</span><span class="hljs-punctuation">)</span><br>system<span class="hljs-punctuation">(</span>paste<span class="hljs-punctuation">(</span><span class="hljs-string">&quot;/opt/conda/bin/python3 /home/jovyan/upload/zl_liu/wecomchan.py&quot;</span><span class="hljs-punctuation">,</span> <span class="hljs-string">&quot;&#x27;task p3 successfully completed&#x27;&quot;</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> intern <span class="hljs-operator">=</span> <span class="hljs-built_in">T</span><span class="hljs-punctuation">)</span><br><br>infercnv_obj_p4 <span class="hljs-operator">=</span> infercnv<span class="hljs-operator">::</span>run<span class="hljs-punctuation">(</span>infercnv_obj_p4<span class="hljs-punctuation">,</span><br> cutoff<span class="hljs-operator">=</span><span class="hljs-number">0.1</span><span class="hljs-punctuation">,</span> <span class="hljs-comment"># use 1 for smart-seq, 0.1 for 10x-genomics</span><br> out_dir<span class="hljs-operator">=</span><span class="hljs-string">&quot;p4&quot;</span><span class="hljs-punctuation">,</span> <span class="hljs-comment"># dir is auto-created for storing outputs</span><br> cluster_by_groups<span class="hljs-operator">=</span><span class="hljs-built_in">F</span><span class="hljs-punctuation">,</span> <br> analysis_mode<span class="hljs-operator">=</span><span class="hljs-string">&quot;subclusters&quot;</span><span class="hljs-punctuation">,</span><br> tumor_subcluster_partition_method <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;random_trees&#x27;</span><span class="hljs-punctuation">,</span><br> denoise<span class="hljs-operator">=</span><span class="hljs-literal">TRUE</span><span class="hljs-punctuation">,</span><br> HMM<span class="hljs-operator">=</span><span class="hljs-literal">TRUE</span><span class="hljs-punctuation">,</span><br> num_threads<span class="hljs-operator">=</span><span class="hljs-number">12</span><span class="hljs-punctuation">)</span><br>saveRDS<span class="hljs-punctuation">(</span>infercnv_obj_p4<span class="hljs-punctuation">,</span> <span class="hljs-string">&#x27;infercnv_obj_p4.rds&#x27;</span><span class="hljs-punctuation">)</span><br>system<span class="hljs-punctuation">(</span>paste<span class="hljs-punctuation">(</span><span class="hljs-string">&quot;/opt/conda/bin/python3 /home/jovyan/upload/zl_liu/wecomchan.py&quot;</span><span class="hljs-punctuation">,</span> <span class="hljs-string">&quot;&#x27;task p4 successfully completed&#x27;&quot;</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> intern <span class="hljs-operator">=</span> <span class="hljs-built_in">T</span><span class="hljs-punctuation">)</span><br><br>infercnv_obj_p5 <span class="hljs-operator">=</span> infercnv<span class="hljs-operator">::</span>run<span class="hljs-punctuation">(</span>infercnv_obj_p5<span class="hljs-punctuation">,</span><br> cutoff<span class="hljs-operator">=</span><span class="hljs-number">0.1</span><span class="hljs-punctuation">,</span> <span class="hljs-comment"># use 1 for smart-seq, 0.1 for 10x-genomics</span><br> out_dir<span class="hljs-operator">=</span><span class="hljs-string">&quot;p5&quot;</span><span class="hljs-punctuation">,</span> <span class="hljs-comment"># dir is auto-created for storing outputs</span><br> cluster_by_groups<span class="hljs-operator">=</span><span class="hljs-built_in">F</span><span class="hljs-punctuation">,</span> <br> analysis_mode<span class="hljs-operator">=</span><span class="hljs-string">&quot;subclusters&quot;</span><span class="hljs-punctuation">,</span><br> tumor_subcluster_partition_method <span class="hljs-operator">=</span> <span class="hljs-string">&#x27;random_trees&#x27;</span><span class="hljs-punctuation">,</span><br> denoise<span class="hljs-operator">=</span><span class="hljs-literal">TRUE</span><span class="hljs-punctuation">,</span><br> HMM<span class="hljs-operator">=</span><span class="hljs-literal">TRUE</span><span class="hljs-punctuation">,</span><br> num_threads<span class="hljs-operator">=</span><span class="hljs-number">12</span><span class="hljs-punctuation">)</span><br>saveRDS<span class="hljs-punctuation">(</span>infercnv_obj_p5<span class="hljs-punctuation">,</span> <span class="hljs-string">&#x27;infercnv_obj_p5.rds&#x27;</span><span class="hljs-punctuation">)</span><br>system<span class="hljs-punctuation">(</span>paste<span class="hljs-punctuation">(</span><span class="hljs-string">&quot;/opt/conda/bin/python3 /home/jovyan/upload/zl_liu/wecomchan.py&quot;</span><span class="hljs-punctuation">,</span> <span class="hljs-string">&quot;&#x27;task p5 successfully completed&#x27;&quot;</span><span class="hljs-punctuation">)</span><span class="hljs-punctuation">,</span> intern <span class="hljs-operator">=</span> <span class="hljs-built_in">T</span><span class="hljs-punctuation">)</span><br></code></pre></td></tr></table></figure>
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