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- Click a Single cell counts data node
- Click the QA/QC section of the task menu
- Click Single cell QA/QC
By default, all samples are used to perform QA/QC. You can choose to split the sample and perform QA/QC separately for each sample.
If your Single cell counts data node has been annotated with a gene/transcript annotation, the task will run without a task configuration dialog. However, if you imported a single cell counts matrix without specifying a gene/transcript annotation file, you will be prompted to choose the genome assembly and annotation file by the Single cell QA/QC configuration dialog (Figure 1). Note, it is still possible to run the task without specifying an annotation file. If you choose not to specify an annotation file, the detection of mitochondrial counts will not be possible.
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SubtitleText | Use the Configuration options on the left customize the appearance of plots |
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AnchorName | Configure single cell QA/QC plot |
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High-quality cells can be selected using the Selection card on the right Select & Filter, which is pre-loaded with three the selection rulescriteria, one for each quality metric (Figure 5).
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SubtitleText | The Selection card with three selection rulesSelect & Filter criteria |
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AnchorName | Selection card Select & Filter Single cell QAQC |
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Hovering the mouse over one of the selection rules criteria reveals a histogram showing you the frequency distribution of the respective quality metric. The minimum and maximum thresholds can be adjusted by clicking and dragging the sliders sliders or by typing directly into the text boxes for each selection criteria (Figure 6).
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SubtitleText | Use the sliders and histograms to adjust the selection criteria |
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AnchorName | Selecting cells using sliders |
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Alternatively, Pin histogram to view all of the thresholds can be set by typing directly into the text boxes in each selection rule distributions at one time to determine thresholds with ease (Figure 7).
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SubtitleText | Minimum and maximum thresholds can be set by manually typing into the text boxes |
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AnchorName | Selecting cells using text boxes |
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| Image RemovedPin the histogram to view all of the distributions at one time | AnchorName | Pin histogram |
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Adjusting the selection criteria will select and deselect cells in all three plots simultaneously. Depending on your settings, the deselected points will either be dimmed or gray. The filters are additive. Combining multiple filters will include the intersection of the three filters. The number of cells selected is shown in the figure legend of each plot (Figure 8).
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SubtitleText | The figure legends show 3696/4352 cells with total count <12,000, number of detected features 600 - 2500, and % mitochondrial count <5% are selected in all three plots. Selected cells are blue, filters are additive and deselected cells are dimmed. |
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AnchorName | Single cell QAQC with selection criteria applied |
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To filter the high-quality cells, click the the include selected cells icon in Filter in the Filtering card in the top right of Select & Filter, and click Apply observation filter... (Figure 9).
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SubtitleText | Include the selected points and apply the filter |
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AnchorName | Apply filter |
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Select the input data node for the filtering task and click Select (Figure 10).
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SubtitleText | After the Apply filter button is selected, you will be presented with a preview of your pipeline. You need to select the appropriate data node to apply the filtering to |
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AnchorName | Select input data node for filtering |
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A new data node, Filtered counts, will be generated under the Analyses tab (Figure 11).
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SubtitleText | Filter cells task runs from the Single cell QA/QC report |
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AnchorName | Output of Single cell QA/QC |
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Double click the Filtered counts data node to view the task report. The report includes a summary of the count distribution across all features for each sample; a detailed breakdown of the number of cells included in the filter for each sample; and the minimum and maximum values for each quality metric (expressed genes, total counts, etc) across the included cells for each sample (Figure 12).
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SubtitleText | Filtered counts task report |
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AnchorName | Filter counts report |
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