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Each row of the spreadsheet (Figure 1) corresponds to a single sample. The first column is the names of the .idat files and the remaining columns are the array probes. The table values are β-values, which correspond to the percentage methylation at each site. A β-value is calculated as the ratio of methylated probe intensity over the overall intensity at each site (the overall intensity is the sum of methylated and unmethylated probe intensities). 

 

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SubtitleTextSpreadsheet after .idat file import: samples on rows (Sample IDs are based on file names), probes on columns, cell values are functionally normalized beta values (default settings)
AnchorNametop level spreadsheet

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The Create categorical attribute dialog (Figure 3) allows us to create groups for a categorical attribute. By default, two groups are created, but additional groups can be added. 

 

 

 

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SubtitleTextCreate Categorical Attribute is used to define new groups and assign samples to them. The name of new column (attribute) is specified in the Attribute name field, group labels (attribute levels) are specified in the Group Name fields. To assign samples to groups, use drag and drop (to select more than one, use Ctrl or Shift buttons)
AnchorNameassigning samples to groups

 

  • Select New Group twice to add two additional groups
  • Set Attribute name: to hPSC 
  • Rename the four groups PrimedNaiveshPOU5F1, and shNANOG
  • Drag and drop the samples from the Unassigned list to their groups as listed in the table below

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There should now be four groups with two samples in each group (Figure 4).

 

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SubtitleTexthPSC attribute with four groups - Primed, Naive, shPOU5F1, and shNANOG - of two samples per group
AnchorNamehPSC attribute

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A new column as been added to spreadsheet 1 (Differential Methylation Analysis) with the experimental group of each sample (Figure 5)

 

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SubtitleTextAdded experimental group as a sample attribute
AnchorNameSpreadsheet with added attribute

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