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Based on pre-alignment QA/QC, we need to trim low quality bases from the 3' end of reads. 

  • Click the Unaligned reads data node
  • Click Pre-alignment tools in the task menu
  • Click Trim bases (Figure 1) 

Figure 1. Invoking the Trim bases task

By default, Trim bases removes bases starting at the 3' end and continuing until it finds a base pair call with a Phred score of equal to or greater than 20. Hover over the  for additional information on any task option (Figure 2). 

  • Click Finish to run Trim bases with default settings

 

Figure 2. Viewing additional information about the Quality score trimming method

The Trim bases task will generate a new data node, Trimmed reads. While tasks have been queued or are in progress they have a lighter color. Any output nodes that the task will generate, such as the Trimmed reads data node from the Trim bases task, are also displayed in a lighter color until the task completes. Once the task begins running, a progress bar is displayed on the task node (Figure 3). 

   

Figure 3. Task and Data nodes that have been queued or are in progress are shown in a lighter color than completed tasks.
We can view the task report for Trim bases by selecting either the Trim bases task node or the Trimmed reads data node and choosing Task report from the task menu. 

  • Select the Trimmed reads data node
  • Select Task report from the task menu

The report shows the percentage of trimmed reads and reads removed in a spreadsheet (Figure 4) and a graph (Figure 5).  

 

Figure 4. Spreadsheet showing results of the Trim bases task
Figure 5. Bar chart showing results of Trim bases task

The average quality score for each base call at each position for each sample is also shown as a graph (Figure 6).

 

Figure 6. Updated base call quality graph after Trim bases task

The results are fairly consistent across samples with ~65% of reads untrimmed, ~30% trimmed, and ~5% removed for each. 

 

 

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