QIAseq Pro Fastq to Somatic CNV Control

The QIAseq Pro Fastq to Somatic CNV Control template workflow produces coverage tables that can be used as controls for copy number variant detection.

Use the workflow to generate coverage tables for the QIAseq Pro Fastq to Annotated Somatic Variants (QIAseq Pro Fastq to Annotated Somatic Variants) template workflow.

QIAseq Fastq to Somatic CNV Control can be found at:

        Template Workflows | LightSpeed Workflows (Image lightspeed_wf_folder_open_16_n_p) | QIAseq workflows (Image qia_ls_folder_open_16_n_p) | QIAseq Targeted DNA Pro (Image qia_ls_pro_folder_open_16_n_p) | QIAseq Pro Fastq to Somatic CNV Control (Image qia_ls_pro_wf_16_n_p)

If you are connected to a CLC Server via your Workbench, you will be asked where you would like to run the analysis. We recommend that you run the analysis on a CLC Server when possible.

In the first wizard step, select a Reference Data Set (figure 6.21).

If you have not downloaded the Reference Data Set yet, the dialog will suggest the relevant data set and offer the opportunity to download it using the Download to Workbench button.

This workflow has been set up to process data generated with QIAseq Targeted DNA Pro panels, and it is important to choose the right reference data to get the reads correctly processed.

The off-the-shelf QIAseq Targeted DNA Pro panels are available in the QIAseq DNA Pro Panels hg38 reference data set. If you have not downloaded the Reference Data Set yet, the dialog will offer the opportunity to download it using the Download to Workbench button.

If the QIAseq DNA Pro Panels hg38 reference data set does not contain the needed primers and target regions, a custom reference data set can be created, see http://resources.qiagenbioinformatics.com/manuals/clcgenomicsworkbench/current/index.php?manual=Custom_Sets.html.

The reference data set for QIAseq Targeted DNA panels should not be used with this workflow. The differences in read structure will for example prevent primers from being correctly trimmed.

Image lis_workflow_qia_pro_cnv_somatic_step1
Figure 6.21: Select a reference data set.

In the LightSpeed Fastq to Germline Variants wizard step (figure 6.22) you have the following options:

Image lis_workflow_qia_pro_cnv_somatic_step2
Figure 6.22: Select fastq files.

In the next dialog (figure 6.23), specify the relevant target regions from the drop down list.

Image lis_workflow_qia_pro_cnv_somatic_step3
Figure 6.23: Select target regions.

Repeat the selection of the appropriate track for target primers in the subsequent dialog (figure 6.24).

Image lis_workflow_qia_pro_cnv_somatic_step4
Figure 6.24: Select target primers.

In the QC for Target Sequencing wizard step, define the threshold for minimum coverage (figure 6.25). This threshold is important because it is used in the quality control section of the sample report. In the later wizard step for Create Sample Report, you will be able to adjust the percent bases in the target regions that should meet this threshold.

Image lis_workflow_qia_pro_cnv_somatic_step5
Figure 6.25: Set the coverage threshold. This threshold is used in the quality control section of the sample report.

In the Create Sample Report wizard step, select relevant summary items and specify thresholds for quality control (figure 6.26). Summary items, thresholds and an indication of whether specified thresholds were met, will be shown in the quality control section of the sample report. The default summary items are appropriate for many data sets, but may need to be adjusted.

To add more summary items, press Add..., choose the report type LightSpeed fastq to somatic variants or QC for targeted sequencing and select summary items as appropriate.

Image lis_workflow_qia_pro_cnv_somatic_create_sample_report
Figure 6.26: Specify summary items. These will be shown in the quality control section of the sample report.

In the final wizard step, choose to Save the results of the workflow and specify a location in the Navigation Area before clicking Finish.



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